CB Catalog Biología

2013 Cell biology product and resource guide
Problem? Solved.
By phone, online, or on-site, were here to quickly solve your challenges, no matter how great. Get award-winning support for every issue, every time.
Expert application scientists ready to answer your specific questions Field support in more than 60 countries Comprehensive website with easy access to manuals, protocols & technical documents
Let us help at lifetechnologies.com/support
Its a dynamic time to be working in life sciences.

We know that every day, you keep pressing toward scientific breakthroughs in spite of increased uncertainty, shifting technology, and decreased funding. Your work is already complex enough. But now more than ever, being at the forefront of discovery means navigating through change.
And we are there to support you at every turn.

Thats why we are making a few adjustments on our end to help simplify things on yours. Were assessing how we do thingsimproving where we need it, and continuing to do what already works. Weve made our website more intuitive and straightforward so you can easily determine what products you need and find them all in one place. We are creating new ways to offer products that are priced to meet your budget but dont compromise quality. Were ensuring that our people are highly skilled, readily available, and understand the many facets of your research, so you get answers quicklythe first time, every time. We are making sure our world-class support team members operate more like trusted colleaguesand less like representativesto walk you through solutions step by step, with simple answers and calming reassurance.
Most importantly, we are keeping our focus on making innovation brilliantly simple to improve life in your lab. So you can keep driving toward discoveries that improve life for all of us.
We are here to support you at every turn. Let us know how we can do that at facebook.com/lifetechnologies
Cell biology resources
2013 Life Technologies
Cell biology product and resource guide

Cell biology products
Comprising trusted and familiar Gibco, Invitrogen, Molecular Probes, and Novex products, the Life Technologies cell biology portfolio offers published, high-performance, scalable solutions across the breadth of cell biology applications. The book begins with overviews of five prominent cell biology research areas: neurobiology, immunology, cancer, stem cell biology, and plant cell biology. Using published data, weve also constructed pathway maps for some of these research areas that you may consult as you plan your research.
Cell culture and cells (Chapter 2) Sample preparation

Recovery Freezing Medium, page 40 Gibco RPMI Media 1640, page 41 Gibco Minimal Essential Medium (MEM), page 41 GlutaMAX media, page 41 Opti-MEM reduced-serum medium, page 41
Cell analysis (Chapter 3)

Dynabeads cell isolation technology, page 76 Dynabeads CD3/CD28 technology, page 79
Key technologies
Expi293 Expression System, page 42 FreeStyle MAX CHO Expression system, page 42 AlgiMatrix 3D cell culture system, page 49 Geltrex LDEV-free matrix products, page 49 AmnioMAX media, page 51 MarrowMAX bone marrow medium, page 51 PB-MAX Karyotyping Medium, page 51 KaryoMAX medium, page 51 TrypLE reagent, page 52
Click-iT EdU cell proliferation assay, page 82 Premo FUCCI Cell Cycle Sensor, page 83 CellROX Deep Red Reagent, page 86 CellLight fluorescent protein constructs, page 87 Qtracker Cell Labeling, page 95 FluoroMyelin Green fluorescent myelin stain, page 95 Molecular Probes antibodies for multicolor flow Qdot nanocrystals for flow cytometry, page 105 Zenon antibody labeling, page 107 APEX antibody labeling, page 107
cytometry, page 102
Essentials
page 41 Advanced Gibco media: GlutaMAX, OptiMEM, page 41 Gibco FBS, page 43 Geneticin selection antibiotic, page 46 Puromycin seletion antibiotic, page 46 Zeocin selection antibiotic, page 46 Gibco growth factors, page 47 B-27 Supplement, serum substitute for neural cell culture, page 72 Neurobasal Media, basal media, page 48 KnockOut Serum Replacement, for pluripotent stem cells, page 60
Classical Gibco media: DMEM, RPMI, and MEM,
DynaMag magnets, page 80 HulaMixer Sample Mixer, page 80 LIVE/DEAD Viability/Cytotoxicity Kits, page 81 CellEvent Caspase-3/7 Green Detection Reagent, pHrodo indicators, page 87 Alexa Fluor phalloidins, page 90 MitoTracker dyes, page 91 JC-1 dye, page 91 LysoTracker dyes, page 92 BODIPY FL C5-ceramide, page 93 TO-PRO-3, page 94 Calcein, AM, page 95 CellTracker dyes, page 95 Fluorescent dextrans, page 95 Alexa Fluor conjugated secondary antibodies and
streptavidin, page 106 page 85
Detection instruments
Countess Automated Cell Counter, page 53 Neon Transfection System, page 55 Tali Image-Based Cytometer, page 55 FLoid Cell Imaging Station, page 55 Attune Acoustic Focusing Cytometer, page 55
Tali Image-Based Cytometer, page 99 FLoid Cell Imaging Station, page 101 Attune Acoustic Focusing Cytometer, page 100
2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.
Chapter 1 contains tools you may find useful to choose products and interpret your data.
Cell biology products
The rest of the guide is summarized in the chart belowan organized snapshot of the most popular of our reliable solutions, backed by nearly 3 million publications. For complete product information, refer to the chapter/page numbers noted.
Protein analysis (Chapter 4)

Dynabeads immunoprecipitation, page 129 Novex organelle isolation kits, page 132 Novex protein A/G purification, page 130 Novex cell lysis reagents, page 133
Protein expression
Champion pET Expression System, page 119 PichiaPink Yeast Expression System, page 118 GeneArt Algae Engineering Kit, page 120 GeneArt Gene Synthesis, page 121 Lipofectamine LTX Transfection Reagent, page 123 Lipofectamine RNAiMAX Reagent, page 123 Lipofectamine 2000 Transfection Reagent, page 123 Silencer Select siRNA, page 126 Stealth Select siRNA and Silencer siRNA, page 126 mirVana miRNA Mimics and Inhibitors, page 128
Protein analysis
page 141
Bolt and NuPAGE protein gels, page 136 Novex Sharp and SeeBlue Plus 2 Pre-Stained Standard, SYPRO Ruby Protein Gel Stain, page 142 Novex ABfinity antibodies, page 147 Novex multiplex immunoassays for Luminex platforms, page 153 Novex magnetic multiplex immunoassay for Luminex, page 154
Protein expression
Ambion Pre-miR Precursors and Anti-miR Inhibitors,

page 128 Lipofectamine reagents, page 123
Protein analysis
Novex pour-your-own gel essentials, page 136 Novex Tris-glycine gels, page 136 SeeBlue Pre-Stained Standard, page 141 SimplyBlue SafeStain, page 142 Novex Antibody Pair kits, page 150 Novex ELISA kits, page 150
Qubit Fluorometer, page 134 Neon Transfection System, page 124 POROS analytical chromatography, page 131 ZOOM IEF Fractionator, page 132
XCell SureLock Protein Electrophoresis Cells, page 139 iBlot 7-Minute Blotting System, page 143 BenchPro 4100 Western Processing System, page 144 MAGPIX System, page 155
lifetechnologies.com
0 1 2 3 4
Product type Research use Class I - IVD
Cell biology research and pathways
Cell culture and cells
Cell analysis
Protein analysis
The products listed in this document have the following product uses. Product Use For Research Use Only. Not for use in diagnostic procedures. In vitro Diagnostic Use For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into human or animals. OR For Research Use or Non-Commercial Manufacturing of Cell-Based Products for Clinical Research. Caution: Not intended for direct administration into humans or animals. For human ex vivo tissue and cell culture processing applications: CAUTION: When used as a medical device, Federal Law restricts this device to sale by or on the order of a physician.
CTS brand research use
Class II medical device
Unless otherwise indicated, the products included in this catalog are For Research Use Only. Not for use in diagnostic procedures.
Cell biology research and pathways Cell biology research and pathways
Research areasThis section provides an overview for investigators in a specific discipline to see the broad menu of products, services, and tools Life Technologies offers for their research and tools. Neurobiology research Immunology research Cancer research Stem cell research Plant cell biology research Cellular signaling pathwaysRefer to these publication-supported representations of several important signaling pathways. AKT pathway MAPK pathway JAK/STAT pathway 22 24 26 8 14 16 18 20
Neurodegeneration 28 Custom servicesTake advantage of these terrific service offerings to customize your tools. Gibco custom media services Custom Novex multiplex assays for Luminex technology Custom biology services Website resourcesAll of these resources are available at a click, including resources for learning techniques, finding the right cell line by application, downloading protocols, stain and antibody selection guides, and mobile apps for the scientist on the go. Gibco Cell Culture Basics Cell Line Database Protocol Database The Molecular Probes Handbook online
30 32 33
34 34 35 35 36 36 36 37 37
Fluorescence SpectraViewer Mobile applications and widgets Virtual Cell Staining Tool Antibody and immunoassay selection guides Customer and technical support
Neurobiology research
Life Technologies offers a range of cells and cell culture tools as well as cellular analysis assays and reagents for research on neural primary and stem cells. Our products encompass all major neural cell types and support a multitude of applications from basic research to therapeutic discovery. Select the right tools for your neuroscience research at lifetechnologies.com/neuro.
Cells and culture tools for neurobiology

Neural primary and stem cells Gibco offers a wide array of highly pure, ready-to-use primary and stem cells for neural research that have high viability and superior performance. For more details on cells available, visit lifetechnologies.com/ neuroculture.
Rat cortex and hippocampus neuronal cells Rat primary cortical astrocytes Human astrocytes
Stem cells: Human neural stem cells Rat fetal neural stem cells Rat glial precursor cells Human dopaminergic precursor cells (available as custom order only) Media and supplements Selecting the right media system for your neural cell culture is important to ensure experimental consistency, reproducibility, and relevancy. For over twenty years, neuroscience researchers have relied on Gibco products to ensure confidence in results. Cited in over 11,000 publications spanning more than 50 cell types and an expansive list of disease models, Gibco neural cell culture products deliver the versatility required from media systems today. Choose from supplements optimized to support a variety of applications, basal media designed for key cell types, and quality primary and stem cells. To learn more, visit lifetechnologies.com/neuroculture. To build the optimal media system for your neural cell culture needs, explore our product offering by cell type, or browse our products designed for specific customer applications like electrophysiology or insulin studies, in the tables on pages 9 and 10.
Neural primary cells:
Primary rat hippocampus neurons
Human neural stem cells

Tools for primary neurons.
Supplement B-27 Serum Free Supplement Primary neurons Application Growth and maintenance of neurons Basal media Neurobasal Medium Neurobasal-A Medium DMEM media Optimized cell type Optimized for prenatal and fetal neurons Optimized for postnatal and adult brain neurons Growth of a spectrum of mammalian cell lines Cells Primary rat cortex neurons Primary rat hippocampus neurons Cell source Isolated from day-18 Fisher 344 rat embryos
Isolated from day-18 Fisher 344 rat embryos
G-5 Supplement Glial cells
Growth of glial cell lines
G-5 Supplement Astrocytes
Astrocyte Medium
Growth of primary and serial tumor lines of astrocytic phenotype Growth of primary human and rat astrocytes
Rat primary cortical astrocytes
Isolated from cortices of SpragueDawley rats at embryonic day 18 (E19) Normal human cells derived from human brain tissue
Human astrocytes
Tools for neural stem cells and progenitor cells.

Supplement B-27 Supplement, minus Vitamin A Application Proliferation of stem cells Basal media StemPro NSC SFM Optimized cell type Human neural stem cells Cells Human neural stem cells Cell source Derived from H9 (WA09) human embryonic stem cells Isolated from the cortex of SpragueDawley E14 rats
Neural stem cells
B-27 Supplement, XenoFree N-2 Supplement
Differentiation of stem cells Serum substitute for embryonic neural cells Growth and expansion of human and rat neural stem cells (NSCs) Serum substitute for CNS progenitor cells Growth and expansion of progenitor cells
KnockOut DMEM/ F-12 Medium
Neural stem cells
Rat fetal neural stem cells
StemPro Neural Supplement
Progenitor cells
N-2 Supplement
KnockOut DMEM/ F-12 Medium
Neural stem cells
Rat glial precursor cells (rGPCs) Human dopaminergic precursor cells*
Isolated from newborn SpragueDawley rat cortex Derived from H9 human embryonic stem cells (ESCs)
StemPro Neural Supplement
* Available as custom order.

Tools optimized for specialty applications.
Supplement B-27 Supplement, minus AO (antioxidants) Application Studies of oxidative stress/damage, apoptosis, or where free radical damage to neurons occurs Studies of insulin secretion or insulin receptors Electrophysiology studies Growth and expansion of retinal ganglion cells Application Receptor studies, downstream purification studies, or other processes where the presence of phenol red is undesired Manipulation of neurons at ambient CO2 for at least 48 hr while retaining viability Optimized cell type Prenatal and fetal neurons Optimized for postnatal and adult brain neurons Optimized for postnatal neurons Optimized for embryonic neurons
B-27 Supplement, minus insulin B-27 Electrophysiology Kit B-27 Supplement, custom Basal media Neurobasal Medium without phenol red
Neurobasal-A Medium without phenol red Hibernate-A Medium Hibernate -E Medium
Serum-free supplements
B-27 Supplementsoptimized serum substitute for neural cell culture available exclusively from Life N-2 Supplementserum substitute for embryonic neural and central nervous system progenitor cells G-5 Supplementchemically defined, serum-free supplement for growth of glial cell lines StemPro Neural Supplementserum-free supplement formulated for the growth and expansion of human Gibco seradeliver consistent cell growth over time and passages; learn more at lifetechnologies.com/
cellculture and rat NSCs and progenitor cells Technologies; learn more at lifetechnologies.com/b27
Supplement comparison.
Standard for mammalian cells Product Lot-to-lot consistency cGMP manufacturing Performance testing Compatible cell types Formulations offered Learn more Gibco sera Highest grade only Yes Non-neural cell line proliferation Neural and non-neural cell lines Qualified and certified lifetechnologies.com/cellculture Industry standard for neuronal cell culture B-27 Serum Free Supplement Yes Yes Primary neuron cell survival Human and animal primary neurons and glia Minus antioxidants, minus insulin, or electrophysiology lifetechnologies.com/b27 Optimized for neuronal stem cells StemPro Neural Supplement & B-27 Supplement, XenoFree Yes Yes Proliferation and differentiation of human NSCs Human and animal NSCs Minus vitamin A and XenoFree lifetechnologies.com/NSC
10

Basal media
Neurobasal mediasupport growth of neurons when combined with B-27,

N-2, or G-5 supplements and designed to inhibit glial proliferation; learn more at lifetechnologies.com/neurobasal KnockOut DMEM/F-12 Mediumoptimized for growth of undifferentiated stem cells; supports growth of NSCs when used with StemPro Neural Supplement StemPro NSC SFMserum-free medium formulated for growth and expansion of human NSCs derived from either ESCs or fetal tissue; kit includes KnockOut DMEM/F-12 medium, StemPro Neural Supplement, and growth factors Gibco DMEMClassic formulation used to culture glial cells (astrocytes and oligodendrocytes) Cell substrates
Natural mouse lamininimportant for cell attachment, spreading, and neurite Fibronectinmodulates cell adhesion, growth, migration, and differentiation as a CELLstart CTS Substratefully defined, xeno-free substrate for stem cell
growth and expansion; maintains multipotency of human NSCs in StemPro NSC SFM Enzymatic passaging component of the extracellular matrix outgrowth
StemPro Accutase Cell Dissociation Reagentdesigned for gentle and efficient

detachment of neural stem cells, progenitor cells (including glial), and primary neural cells Cryopreservation
Synth-a-Freeze Cryopreservation Mediuma defined liquid cryopreservation Hibernate mediaideal for manipulation of neurons at ambient CO2 and as preservation media for viable brain tissue for up to a month at 4C medium for human keratinocytes, ESCs, NSCs, and mesenchymal stem cells
11

Cellular analysis assays and reagents for neurobiology
Cell viability assays We have a range of cell viability assays to assess the health of your neural cells. View more information and compare cell viability assays at lifetechnologies.com/cellhealthneuro.
Types of cell viability assays.

Fast resazurin-based assays Product Assay type Mechanism of action Format Incubation time Readout Instrument compatibility For more info, visit: PrestoBlue Cell Viability Reagent Single parameter Metabolic activity Live >10 min Absorbance or fluorescence Microplate reader lifetechnologies.com/ prestoblue Quantitative measurement of DNA content CyQUANT Cell Proliferation Assay Single parameter DNA quantification Live 3060 min Fluorescence Microplate reader, microscopy lifetechnologies.com/ cyquantdirect Quantitative measurement of neurite outgrowth Neurite Outgrowth Staining Kit Multiplexed Esterase activity, membrane integrity Live or endpoint 30 min Fluorescence Microplate reader, microscopy lifetechnologies.com/ neuriteoutgrowth Simultaneous quantification of live and dead cells LIVE/DEAD cell viability assays Multiplexed Esterase activity, membrane integrity Endpoint 1545 min Fluorescence Flow cytometry, microplate reader, microscopy lifetechnologies.com/ livedead
Fluorescent probes for neuronal physiology Choose from a diverse selection of fluorescent ion indicators and probes for calcium, sodium, zinc, potassium, magnesium, pH, membrane potential, and synaptic function from the industry leader in fluorescent technology.
FluxOR Potassium Ion Channel Assayan optically based, homogeneous assay for high-throughput
screening measurements of potassium ion channel and transporter activities; learn more at lifetechnologies.com/fluxor Fluo-4 Direct Calcium Assay Kitadvanced formulation that suppresses background fluorescence generated from media with negligible impact on the cellular fluorescence, allowing the assay to be run in a simple addition-only format in the presence of serum-containing media; learn more at lifetechnologies.com/fluo4direct Voltage sensor probesthese probes use a fluorescence resonance energy transfer-based voltage-sensing assay technology to measure changes in cell membrane potential; learn more at lifetechnologies.com/vsp FM probes for synaptic functionthese probes are ideal for investigating the mechanisms of synaptic vesicle activity; learn more at lifetechnologies.com/fmprobes Probes for neuronal tracing and anatomy Choose from an extensive range of fluorescent and biotinylated tracers for retrograde and anterograde tracing, fluorescent histological stains, and ligands for receptors, ion channels, and ion carriers. Visit lifetechnologies.com/neuroprobes for a full list of probes for neuronal tracing and anatomy.
12

Detection of neuro-related proteins Tools for neuro-related protein detection.
Novex product Qualitative Western blotting Analytes per well Hands-on time Dynamic range For more info visit One 23 days Low Western products: lifetechnologies.com/westerndetection Antibodies: lifetechnologies.com/abfinity lifetechnologies.com/antibodies Industry standard ELISA One 35 hours Medium lifetechnologies.com/neuroelisa Multiplexable Multiplex assay using Luminex platform Up to 50 (proteins) 45 hours High lifetechnologies.com/luminex
Cellular expression analysis BacMam technology BacMam technology allows simple, safe, efficient delivery and expression of genes of interest in mammalian cells, including neurons. For more information about BacMam technology, visit lifetechnologies.com/bacmam. Cellular and pathway analysis Target-based assays Select from our broad range of validated cell-based assays for key neural targets, including G-protein BacMam reagentkinases, construction coupled receptors, and ion channels. Visit lifetechnologies.com/discoveryassays. Custom assay development Let us create and validate a custom assay that meets your defined requirements. Visit lifetechnologies.com/custom.
Target gene tranfer plasmid
Clone into baculovirus DNA
Transfect S9 insect cells
Collect BacMam virus
Assay development and screeening
Use BacMam stocks
Transduce mammalian cells
Con gure assays
Perform screen or assay
BacMam workflow. Just add the BacMam reagent to your cells for 24 hours, treat with an enhancer (recommended for certain cell types including neurons), wash, incubate overnight, and perform the assay.
13
Immunology research
Immunology research
At Life Technologies, we understand the importance of using the highest-quality and most sensitive reagents to dissect the intricacies associated with immunology research. Our extensive portfolio of products can be used across all stages of T and B cell development and activation, from isolation to stimulation and from expansion to analysis.
Isolate: Dynal magnetic cell separation technologies yield pure, Stimulate: Our validated reagents allow better control using state-of-the-art
expansion technology and Gibco cytokines with consistently high bioactivity. Ambion siRNA technology is ideal to knock down key genes and proteins involved in immunology processes. Analyze: We offer products for flow cytometry, imaging microscopy, western blot analysis, and ELISA as well as Novex multiplex assays for protein quantification using Luminex xMAP technology. Check out our immunology-related signaling pathways on pages 15 and 22-29. These include: uncontaminated cells without columns.
The inflammatory response The AKT pathway The MAPK pathway
The JAK/STAT pathway Neurodegeneration
Find a comprehensive selection of our immunology products at lifetechnologies.com/immunology.
Below is a quick guide to the sections where you can learn more about immunology products and solutions from Life Technologies.
Isolate
Reagents
Dynabeads magnetic cell separation products for: Human cells Mouse cells Other cell types Isolation of your own antibody cells CD4+ cell counting kits Primary antibodies for: Human immunology Mouse immunology Rat immunology
Stimulate
Specialty media for immunology research: Gibco OpTmizer Cell Expansion Serum-Free Medium Gibco Serum-Free AIM V Medium Gibco Macrophage Serum-Free Medium Serum-free medium for hematopoietic stem cell culture Cytogenetic media Recombinant growth factors and cytokines
Analyze
Primary antibodies for: Human immunology Mouse immunology Rat immunology Phosphorylation sitespecific primary antibodies Secondary detection reagents Antibody pairs
Assays and kits
Positive isolation Negative isolation Depletion
Novex multiplex assays for the Luminex platform ELISA kits Cell viability kits Cell proliferation kits Dynabeads technology for mouse and human T cell activation and expansion siRNA Click chemistry Luminex xMAP technology CellLight reagents (BacMam technology) ABfinity recombinant monoclonal antibody technology Attune Acoustic Focusing Cytometer FLoid Cell Imaging Station Tali Image-Based Cytometer MAGPIX System iBlot Western Transfer System Flow cytometry controls and standards Imaging standards and calibration kits Protein labeling kits Chemiluminescence detection systems
Technologies
Instrumentation
Countess Automated Cell Counter
Neon Transfection System
Accessories
Magnets and mixers
14
Inammation
Bacterial products (LPS, peptidoglycans, etc.)
Fibroblast liver Acute phase proteins TNF- IL-1 TGF- PDGF IL-6
PECAM-1, VCAM-1, ICAM-1, P-selectin, E-selectin
TNF, IL-1
Endothelial cell
Immunology research
Fever
IL-8, TNF-
Macrophage
IL-1, IL-6, IL-8, IL-10, IL-12, IL-15, IFN-, IFN-, TNF-, MCP-1, MIP-1
M-CSF, TNF-, GM-CSF, G-CSF, IL-1, IL-6, IL-11, IL-12

Hematopoiesis
Cardiovascular pathology
Hi st am in e
Neutrophil elastase Neutrophil Respiratory burst Reactive oxygen species
IL-18 IL-3, IL-4, IL-10, IL-13, IFN-, TNF- IL-3, IL-6, IL-7, GM-CSF IL-3, IL-4, IL-10
Resting T cell
Resting B cell
IL-10, IL-14, IL-4, IL-6
Mast cell
IL-12, IL-15
Allergy anaphylaxis Innate immunity
IL-3, IL-4, IL-5
Activated LFA-1, VLA-4, T cell

IFN- IL-6 Mac-1 IL-2
Eotaxin
Eosinophil
Activated B cell
IL-23 TGF-, IL-2, IL-4,IL-5, IL-13, IFN-
Lung epithelium
IL-2
Viral infection, tumor surveillance
NK cell Th17/22 Th1/Th2 CTL
Plasma cell
IgE IgM/G/A
Humoral immunity Allergy anaphylaxis
Th9
IL-2, IFN-, IL-4
Fibrosis Autoimmunity Tissue repair, protozoan immunity
Viral infection, tumor surveillance
IL-17A, IL-17F, IL-22, IL-6, GM-CSF
The inflammatory response in immunology.
15
Cancer research
Cancer research
At Life Technologies, we are dedicated to advancing cancer research and the pursuit of a world without cancera goal we share with you, the research and medical community. We are here to provide you with the instruments, reagents, and technologies for cancer genomic and cancer cell analysis applications to help translate your ideas from basic cancer research to future clinical applications.
Translating genomic discoveries from the bench to the bedside and beyond
The key to fighting cancer through better therapeutics depends on a better understanding of the basic biology of this disease. A major challenge in cancer research is unraveling the causes of cancer on a molecular level. Because most of the genomic mutations or alterations in cancer impact the way that cells communicate with other cells, a solid understanding of both genomic and cellular mechanisms is needed to accelerate cancer research. At Life Technologies, we are committed to providing the widest technology selection with the highest quality at every budget to help address challenges in cancer genomics and cancer cellular analysis applications.
Cancer Cellular Analysis Applications
Cancer Genomics Applications

Targeted Sequencing & Gene Panels
Cell Viability & Proliferation Cancer Cell Imaging Cell Signaling & Pathway Analysis Whole Genome & Whole Exome Sequencing
Gene Expression by Sequencing
Cancer Stem Cells
Cancer Cell Growth & Manipulation
Genotyping Assays-Somatic Germline Whole Transcriptome
Gene ExpressionFocused panels & assays
Epigenetics
Cancer Immunotherapy
Observe, follow, and probe deeper into cellular networks
Identify changes in the genomes of tumors that drive cancer progression
Overview of key cancer cellular analysis and genomics applications.
16

Cancer cellular analysis and genomic applications
Cancer cellular analysis involves understanding complex defects in communication between cells that alter normal programs of proliferation, transcription, growth, migration, differentiation, and death. Further advances in understanding altered cancer pathways and their interconnections may accelerate the development of targeted molecular therapies allowing improvements in cancer diagnosis and treatment. The right mix of tools and technologies are needed to observe, follow, and probe deeper into cancer cellular networks. With expertise that spans from cell culture, RNAi, and gene expression analysis to innovative products for rapid protein detection and cellular imaging, we have affordable, high-quality solutions designed to help you reach your cancer research goals. Learn more about the tools we offer for these applications below. Cell viability, proliferation, and function Evaluate the hallmarks of cancer with Molecular Probes assays validated on multiple instrument platforms, including microscopes, flow cytometers, microplate readers, and high-content screening instruments. To learn more, visit lifetechnologies.com/cellviability. Cellular imaging Visualize intracellular changes in cancer cells right on your benchtop using Molecular Probes assays and the FLoid Cell Imaging Station. To learn more, visit lifetechnologies.com/floid. Image cell cycle progression in live cells with the Premo FUCCI Cell Cycle Sensor. To learn more, visit lifetechnologies.com/ premochameleon. Cell signaling pathways Analyze how cancer cell signaling pathways are disrupted in cancer by accurate quantitation of intracellular and extracellular proteins using protein analysis tools designed for single or multiplex cancer biomarker analysis. To learn more, visit lifetechnologies.com/immunoassay and lifetechnologies.com/abfinity. Cell growth and manipulation Foster the right cells and culture environments to decipher the cancer cell signaling pathways influenced by altered genes. To learn more, visit lifetechnologies.com/cellculture. Identify genes and processes regulated by your gene or pathway of interest through the power of RNAi. To learn more, visit lifetechnologies.com/rnai. Use targeted editing of the genome to precisely evaluate cancer phenotypes through target validation and cellline optimization. To learn more, visit lifetechnologies.com/geneart. Rare cell event analysis Identify, track, and analyze cancer cell signaling events in individual cancer cells using flow cytometry to study cell proliferation, the cell cycle, apoptosis, and immunophenotyping and to detect rare cellular events such as the presence of circulating endothelial cells. To learn more, visit lifetechnologies.com/attune. Cancer stem cells Identify quiescent cancer stem cells and determine their role in metastasis and therapeutic resistance. To learn more, visit lifetechnologies.com/cancerstemcells. Cancer immunotherapy Efficiently isolate, activate, and culture various types of immune cells for your cancer immunotherapy research needs. To learn more, visit lifetechnologies.com/ immunotherapyresearch. Drug discovery and development Drive efficiency in drug assay development, high-throughput screening, and metabolic and safety assessment with the support, expertise, and customization to meet your unique cancer research needs. To learn more, visit lifetechnologies.com/drugdiscovery. Genotype-to-phenotype research Determine normal and cancer-related functions of genes, and the implication of variation within those genes, to reach a better understanding of cancer. To learn more, visit lifetechnologies.com/cancergenotype. Cancer epigenetics The initiation and progression of cancer, traditionally seen as a genetic disease, is now realized to involve epigenetic abnormalities along with genetic alterations. Recent advancements in the rapidly evolving field of cancer epigenetics have shown that disruption of various epigenetic mechanisms, including noncoding RNAs such as microRNAs, DNA methylation, histone modifications, and nucleosome positioning, are central to switching normal cells to malignant cells. Life Technologies has developed powerful epigenetic research technologies spanning renewable antibody development, sample preparation, sequencing, quantitative PCR, and functional analysis assays. To learn more, visit lifetechnologies.com/ cancerepigenetics. Renewable epigenetic antibodies Epigenetic regulatory proteins are involved in a wide variety of chronic diseases, including cancer, and are the target of an explosive new field of drug discovery. The ability to properly study most epigenetic changes, which determine the genes that are turned off and on, are determined by the effectiveness of one critical toolantibodies. Life Technologies has partnered with the Structural Genomics Consortium to develop high-quality recombinant antibodies to read, write, and erase epigenetic code. To learn more, visit lifetechnologies .com/epiantibodies. Noncoding RNAs that regulate cancer Discovering how noncoding RNAs regulate gene expression will lead to new approaches to understanding cancer. At Life Technologies, we are developing sensitive methods to uncover new RNAs, solutions to profile and validate microRNAs, and functional analysis tools to help you define the most important noncoding RNAs in cancer. To learn more, visit lifetechnologies.com/cancerrna. Cancer genomics We are committed to accelerating cancer research with highly accurate and sensitive sequencing and real-time PCR systems to quickly find the cancer mutations that matter. To learn more, visit lifetechnologies.com/cancerbiomarkers.
Cancer research
17
Stem cell research

Stem cell research
The discovery in 2006 that human and mouse fibroblasts could be reprogrammed to generate induced pluripotent stem cells (iPSCs) with qualities remarkably similar to embryonic stem cells has created a valuable new source of pluripotent cells for drug discovery, cell therapy, and basic research. Life Technologies products have been an integral part of stem cell research from before the discovery of iPSCs to current breakthroughs. With Gibco stem cell culture products, Invitrogen stem cell reprogramming products, and a wide array of stem cell analysis tools that include Molecular Probes labeling and detection reagents, we have the products to support you at every step of your research.
Stem cell research can be time-consuming finding support shouldnt be

Life Technologies gives you more PSC resources than any other sourceeasy access to more than 70 proven protocols, >2,500 peer-reviewed publications, how-to videos, and webinars from leaders in the stem cell community. And when you have a question, technical support is always available by phone, email, live chat, or in person. We offer resources for areas of stem cell research from culture to reprogramming and tools for stem cell analysis and detection.
Culture4 media options, >20 protocols, and 50 years of innovation Reprogramming3 reprogramming tools, 9 protocols, 4 webinars, and scientists standing by Detection2 iPSC imaging methods, 1 instrument, and >30,000 colleagues
For more information about our resources for stem cell research, go to lifetechnologies.com/pscresources.
Culture
Whether youre studying stem cells for basic research, drug discovery, or therapeutic applications, Life Technologies offers an extensive range of Gibco media, supplements, and reagents to meet all of your stem cell culture needs. You can culture and expand stem cells in cGMP-manufactured serum-free, feeder-free, or xeno-free cell culture systems. Manufactured with superior quality standards, our products offer the consistency and accuracy you need. Find out about all of the options for PSC culture at lifetechnologies.com/culturepsc.
Selecting the right reprogramming tool to generate iPSCs can leave you with questions. And no one has more answers than Life Technologies. From traditional lentivirus methods to nonintegrating technologies like the CytoTune-iPS Sendai Reprogramming Kit and episomal iPSC reprogramming vectors, our products provide more solutions to meet your safety, efficiency, and budgetary needs. Find out about all of the options for reprogramming at lifetechnologies.com/cellreprogram.
Reprogramming
Detection
Validation is critical for iPSC research. Whether you want to quickly confirm that cells are pluripotent or need flexibility in antibody and dye choice, Molecular Probes labeling and detection assays and imaging stations help you get the confirmation you need. And, all of our labeling and detection technologies are backed by our extensive internal and external support resources, including the Molecular Probes online community and the trusted Molecular Probes Handbook. Find out about all of the options for detection at lifetechnologies.com/detectpsc.
18

Culture systems KnockOut Serum Replacementwell-referenced supplement for feeder-dependent growth of PSCs
Stem cell research
StemPro hESC SFMmedium for feeder-free growth of human PSCs that allows optimization of basic fibroblast growth factor concentration
Essential 8 Mediumoptimized 8-component medium for feeder-free growth of human PSCs
KnockOut SR XenoFree Kitfeeder-free and xeno-free medium for human PSCs, with documentation for cell therapy applications
Reprogramming systems
Lentiviral particlesflexible technology for standard reprogramming lifetechnologies.com/lenti
Episomal iPSC reprogramming vectorsnonintegrating, nonviral technology for iPSC generation
CytoTune-iPS Sendai Reprogramming Kitnonintegrating technology for standard and difficult cell types
Detection systems
Alkaline Phosphatase Live Stainallows for a quick check of pluripotency without compromising cell integrity
Antibodiesoptions for specific staining and visualization of stem cell markers with maximum flexibility lifetechnologies.com/antibodies
FLoid Cell Imaging Stationbenchtop instrument for easy visualization of Alkaline Phosphatase Live Stain and antibodies
19
Plant cell biology research

Understanding the structural and functional relationships of plant cells and tissues is critical to plant research. From studying the morphology of plant cells and walls, organelles, and other cellular compartments all the way to complex gene regulation and expression studies, cell tissue analysis techniques and tools are critical not only for basic plant research but also for programs of crop improvement. For example, demonstrating the mechanism of action of a new trait as part of the regulatory package is critical in trait development of new commercial plant varieties. Key techniques such as immunohistochemistry have been routinely used to label diverse plant tissues and organelles including cell walls, vacuoles, chromatin, nuclei, nuclear membranes, and chloroplasts. To empower your research in plant cell and tissue analysis, Life Technologies has a broad portfolio of fluorescent reagents, kits, and instruments specifically designed with your needs in mind.
Molecular Probes reagents including Alexa Fluor dyes, Qdot nanocrystals, Click-iT detection assays, and ProLong Gold antifade reagent are used in a wide range of plants, for a variety of plant research applications. For example:
Molecular Probes fluorescent dyes and probes
Immunohistochemistry Fluorescence in situ hybridization Organelle and cytoskeletal stains Endocytosis studies Transport studies
Glucose metabolism Viability staining Glutathione detection Calcium and other ion imaging
Rubisco localization in maize leaf section. Rubisco was localized using a rabbit anti-rubisco antibody and visualized using the highly cross-adsorbed Alexa Fluor 488 goat antirabbit IgG antibody (Cat. No. A11034). The 2.0 m maize leaf section illustrates the immunolocalization of rubisco in the chloroplasts of the bundle sheath cells surrounding the vascular bundles. The red fluorescence, localized to the mesophyll plastids, is due to background autofluorescence of chlorophyll. Lignin appears dull green and is localized to the xylem of the vascular bundle; cutin appears bright green and is localized to the cuticle outside the epidermis. Image contributed by Todd Jones, DuPont.
Pectin associated with plasmodesmatal pit fields of kiwifruit cells. Pectin, a component of the cell wall matrix and the main constituent of the middle lamella that forms between daughter cell walls, was tagged with an anti-pectin monoclonal antibody, JIM 5. The primary antibody was detected and visualized with Alexa Fluor 488 goat antirat IgG (Cat. No. A11006). The primary antibody was a gift from Dr. Paul Knox, University of Leeds, UK. Image contributed by Paul Sutherland, The Horticulture and Food Research Institute of New Zealand, Ltd., Mt. Albert Research Centre. Ca2+ gradient in elongating lily pollen tube. Top panel: Pseudocolored image of a pollen tube of Lilium longiflorum injected with fura dextran (Cat. No. F3029). The cell continues elongating and clearly shows a Ca2+ gradient. Bottom panel: The same pollen tube after injection with dibromo-BAPTA (Cat. No. D1211) remains healthy but is no longer elongating. Images contributed by Debra Miller, Dale Callaham, David Gross, and Peter Hepler, University of Massachusetts.
20

Flow cytometry
Flow cytometry represents an ideal means for the analysis of both cells and subcellular particles, with a potentially large number of parameters analyzed rapidly, simultaneously, and quantitatively, thereby furnishing statistically exploitable data and allowing accurate and facilitated detection of subpopulations. Since the first report of the use of this technique in plants 40 years ago, flow cytometry has become an essential tool for the understanding of fundamental mechanisms and processes underlying plant growth, development, and function. Key applications include the study of ploidy levels, genome mapping, localization of sex-specific chromosomes, and plant cell and algal studies. Life Technologies offers a wide range of flow cytometric analysis tools including the Attune Acoustic Focusing Cytometer and fluorescent probes. Learn more and see all of our products at lifetechnologies.com/probes.
Featured product: Alexa Fluor dyes
The leading and most trusted fluorescent dyes available today have been used extensively in the plant sciences. In addition to superior performance, youll get an experienced, problem-solving technical support team, over 30,000 published references, application and experimental tips, and protocols to help with experimental planning. Our range of labeling kits, molecules for tracing cell structures, and secondary detection reagents within the Alexa Fluor product line provide superior brightness and photostability, outperforming conventional fluorescent reagents. Learn more at lifetechnologies.com/alexa. For a comprehensive view of all your products and technologies to empower your plant sciences research, visit lifetechnologies.com/plants. A
106
Scatter
B
PI 640 LP BL3-A
106
105
SSC-A
Nuclei
105
104
104
103 103
104
105
106
103 104
105
106
PI 603-48 BL2-A
PI 603-48 BL2-A
700 600 500

4C
1,000 900 800 700 600 500 400 300 200

8C 16C 2C 4C
Count
2C 8C
300 200 100 0 104
16C 32C
Count
105
400
100 0 0 50 100 150
105
200
250
300
PI 603-48 BL2-A
PI 603-48 BL2-A (10^3)
Nuclear holoploid genome sizing analysis of Arabidopsis thaliana leaf tissue homogenates using the Attune Acoustic Focusing Cytometer. (A) Biparametric density plot of side scatter vs. propidium iodide (PI) fluorescence with Scatter gate surrounding the fluorescent nuclei. (B) Biparametric density plot of PI fluorescence (603/48 vs. 640 nm) of Scatter gate population from A. (C) Logarithmic histogram of PI fluorescence (603/48 nm) of Nuclei gate population from B. (D) Linear histogram of PI fluorescence (603/48 nm) of Nuclei gate population from B. 2C, 4C, 8C, 16C, and 32C denote the C-value for the respective peaks.
21
AKT pathway
AKT pathway
The serine-threonine kinase AKT (also known as protein kinase B) is a protein that acts as a central convergence node in a broadly influential signaling network. AKT activation serves as a master switch of these cellular signaling pathways, generating a multitude of intracellular responses through a plethora of downstream targets and interacting partners. Because of its pivotal role in cell signaling, and the consequences of that signaling in diseases ranging from cancer and diabetes to neurodegeneration, AKT is one of the most actively studied kinases in both basic research and drug development. Here we highlight a few of our products for analyzing the AKT signaling pathway and its downstream targets. Life Technologies offers Novex antibodies, ELISAs, Novex assays for the Luminex platform, and growth factors for key targets in the AKT signaling cascade.
Key targets of interest in the AKT pathway

Protein AKT/protein kinase B mTOR GSK-3B MDM2 NF-B Characteristics Master regulator of cell growth and signaling cascade Cell growth regulator Cell cycle regulator Negative regulator of p53 function Transcriptional activator regulating apoptosis Means of regulation Phosphorylation/ dephosphorylation Phosphorylation/ dephosphorylation Phosphorylation/ dephosphorylation Phosphorylation/ dephosphorylation Phosphorylation/ dephosphorylation Research tools (Chapter 4, pages 146-155) Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform
Additional references
Madonna R, Bolli R, Rokosh G et al. (2012) Targeting phosphatidylinositol 3-kinase-Akt through hepatocyte growth factor for cardioprotection. J Cardiovasc Med doi: 10.2459/JCM.0b013e3283542017. Papadimitrakopoulou V (2012) Development of PI3K/AKT/mTOR pathway inhibitors and their application in personalized therapy for non-small-cell lung cancer. J Thorac Oncol doi: 10.1097/JTO.0b013e31825493eb. Wang G, Pan J, Chen SD (2012) Kinases and kinase signaling pathways: potential therapeutic targets in Parkinsons disease. Prog Neurobiol 98(2):207221. Sheppard K, Kinross KM, Solomon B et al. (2012) Targeting PI3 kinase/AKT/mTOR signaling in cancer. Crit Rev Oncog 17(1):6995.
22
Growth factor
AKT pathway
PIP3
-P
PIP2
+P
p85 p110
RTK
mTORC2
GL
PDK1
+P +P
PTEN
PI3K
mTOR
rictor
+P
AKT
+P
-P
+P +P
+P
TSC2 TSC1
Cytoskeletal reorganization
Rapamycin
PRAS40 Rheb
+P
PP2A 4
+P
+P +P
ASK-1 BAD
AR
+P
+P
p21 p27
+P
MDM2
FKBP
mTORC1
GL
mTOR
+P
raptor
GSK-3
+P
CASP9 FOXO
+P
SK6 4E-BP1
+P +P +P +P
CCND1 IKK
+P
Apoptosis
PDCD4
Cell cycle
NFAT IkB
+P
Cell growth
eIF4E
eIF4B RPS6 eIF4A
-CTN
IB NFB
eIF2B
p53 p21 FAS PTEN CIP1
Kinase Phosphatase Receptor tyrosine kinase Transcription factor Translation factor Hormone receptor
Glycogen synthesis Translation initiation

GATA4 NFAT NFB
FOXO FAS-L p27 KIP1
Transcription
Cell proliferation
The AKT pathway.
23
MAPK pathway
MAPK pathway
The mitogen activated protein kinase (MAPK) pathway is central to cellular signal transduction in response to growth factors, mitogens, cytokines, and stress. The MAPK pathway is regulated through a series of reversible phosphorylation events resulting in a variety of cellular activities including growth, differentiation, development, and apoptosis. Here we highlight a few of our products for analyzing the MAPK signaling pathway and its downstream targets. Life Technologies offers Novex antibodies, ELISAs, Novex assays for the Luminex platform, and growth factors for key targets in the MAPK signaling cascade.
Key targets of interest in the MAPK pathway

Protein MAPK ERK JNK Ras Characteristics Regulator of cell growth and signaling cascade Regulator of growth, development, and poliferation Regulator of inflammation, apoptosis, and differentiation GTPase involved in cell proliferation Means of regulation Phosphorylation/ dephosphorylation Phosphorylation/ dephosphorylation Phosphorylation/ dephosphorylation Cleavage of GTP Research tools (Chapter 4, pages 146-155) Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform
Oeztuerk-Winder F, Ventura JJ (2012) The many faces of p38 mitogen-activated protein kinase in progenitor/stem cell differentiation. Biochem J 445(1):110. Gantke T, Sriskantharajah S, Sadowski M et al. (2012) IB kinase regulation of the TPL-2/ERK MAPK pathway. Immunol Rev 246(1):168182. Ppulo H et al. (2012) The mTOR signalling pathway in human cancer. Int J Mol Sci 13(2):18861918. Aksamitiene E, Kiyatkin A, Kholodenko BN (2012) Cross-talk between mitogenic Ras/MAPK and survival PI3K/Akt pathways: a fine balance. Biochem Soc Trans 40(1):139146. Davies C, Tournier C (2012) Exploring the function of the JNK (c-Jun N-terminal kinase) signalling pathway in physiological and pathological processes to design novel therapeutic strategies. Biochem Soc Trans 40:8589.
24
MAPK pathway
The MAPK pathway.
25
JAK/STAT pathway
The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway plays a central role in transducing signals from a wide array of cytokines and growth factors leading to various cellular functions, including proliferation, growth, development, hematopoiesis, and immune responses.
JAK/STAT pathway
Abnormal constitutive activation of the JAK/STAT pathway has been implicated in various cancers and immune disorders. Thus, inhibitors of the JAK/STAT pathway are being sought. Here we highlight a few of our products for analyzing the JAK/STAT signaling pathway and its downstream targets. Life Technologies offers Novex antibodies, ELISAs, Novex assays for the Luminex platform, and growth factors for key targets in the JAK/STAT signaling cascade.
Key targets of interest in the JAK/STAT pathway

Protein JAKs STATs SOCS PIAS Characteristics Initiate signaling upon receptor multimerization Transcription factors Regulator of JAK signaling Regulator of STAT signaling Means of regulation Research tools (Chapter 4, pages 146-155) Phosphorylation/dephosphorylation Novex antibodies, ELISAs, and Novex assays for the Luminex platform Phosphorylation/dephosphorylation, DNA binding Binding of JAK Binding of STAT Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform Novex antibodies, ELISAs, and Novex assays for the Luminex platform
Harrison DA (2012) The Jak/STAT pathway. Cold Spring Harb Perspect Biol 4(3). pii: a011205. doi: 10.1101/cshperspect.a011205. Sansone P, Bromberg J (2012) Targeting the interleukin-6/Jak/stat pathway in human malignancies. J Clin Oncol 30(9):10051014. Wagner KU and Schmidt JW (2011) The two faces of Janus kinases and their respective STATs in mammary gland development and cancer. J Carcinog 10:32. doi: 10.4103/1477-3163.90677. Mohr A, Chatain N, Domoszlai T et al. (2012) Dynamics and non-canonical aspects of JAK/STAT signalling. Eur J Cell Biol 91(67):524532. Tamiya T, Kashiwagi I, Takahashi R et al. (2011) Suppressors of cytokine signaling (SOCS) proteins and JAK/STAT pathways: regulation of T-cell inflammation by SOCS1 and SOCS3. Arterioscler Thromb Vasc Biol 31(5):980985.
26
JAK/STAT pathway
The JAK/STAT pathway.
27
Neurodegeneration
Neurodegeneration
Neuronal death is pivotal to Parkinsons disease (PD) pathogenesis and can be triggered by various cellular mechanisms, including oxidative stress, misfolding of proteins, mitochondrial dysfunction, and proteasome dysfunction. In their pursuit to fully understand the various pathways that lead to neurodegenerative diseases such as PD, researchers are exploring the molecular basis of these mechanisms. Here we highlight just a few of our products for analyzing the key factors and effectors in PD. Life Technologies offers Novex antibodies, ELISAs, Novex assays for the Luminex multiplex assays platform, and growth factors for key targets in studying Parkinsons disease. Some key targets of interest are listed in the table below.
Protein Parkin Characteristics Mitochondrial health Mitochondrial health Leucine-rich kinase Nucleation and stabilization of microtubules Means of regulation Mutation and misfolding Aggregation and mutation Phosphorylation/dephosphorylation Phosphorylation/dephosphorylation/ misfolding Research tools (Chapter 4, pages 146-155) Novex antibodies Novex antibodies, ELISA, and Novex assays for the Luminex platform Novex antibodies, ELISA, and Novex assays for the Luminex platform Novex antibodies, ELISA, and Novex assays for the Luminex platform
-Synuclein
LRRK2 Tau
Protter D, Lang C, Cooper AA (2012) Synuclein and mitochondrial dysfunction: a pathogenic partnership in Parkinsons disease? Parkinsons Dis 2012:829207. Sutachan JJ, Casas Z, Albarracin SL et al. (2012) Cellular and molecular mechanisms of antioxidants in Parkinsons disease. Nutr Neurosci 15(3):120126. Harvey L, Boksa P (2012) Prenatal and postnatal animal models of immune activation: Relevance to a range of neurodevelopmental disorders. Developmental Neurobiology special issue: Neuroimmunology in Development and Disease. Dev Neurobiol doi: 10.1002/dneu.22043. Wang G, Pan J, Chen SD (2012) Kinases and kinase signaling pathways: potential therapeutic targets in Parkinsons disease. Prog Neurobiol 98(2):207221. Rademakers R, Neumann M, Mackenzie IR (2012) Advances in understanding the molecular basis of frontotemporal dementia. Nat Rev Neurol doi: 10.1038/nrneurol.2012.117.
28
Neurodegeneration
Neurodegeneration.
29
Gibco custom media services

Delivering speed, scalability, and quality
Scalability is critical for clinical and biomanufacturing applications. Our tech transfer process helps ensure smooth scale-up as batch size increases.
Small-scale media production when speed matters most
Large-scale media production in our worldclass, validated manufacturing facilities
Choose Gibco MediaExpress for small-scale media production:
Choose Gibco Custom Cell Culture Services for large-scale media production:
Fast turnaround (typically <10 days) Small-scale batches (<200 L, <10 kg) Non-cGMP production 5,000 ft2 pilot plant, Grand Island, NY
Learn more at lifetechnologies.com/mediaexpress.
Industry-leading quality systems and support Large-scale batches (up to 10,000 L)

Learn more at lifetechnologies.com/custommedia.
30

Gibco PD-Direct Bioprocess Services provides media development and optimization services
Gibco PD-Direct services provide contract services leveraging the latest technologies and products from Life Technologies to solve cell-line, media, and process development challenges. PD-Direct services can be your reliable outsourcing partner to expand your capabilities and deliver on-time exceptional results. Choose Gibco PD-Direct Bioprocess Services to partner with technical and industry experts who can assist you every step of the way. Learn more at lifetechnologies.com/bioproduction.
Customize the media format, quality control (QC) testing, and documentation to your needs
Choose a media format and batch size: Liquid: 1 to 10,000 L Powder: 1 kg to 8,000 kg Concentrates: 10 to 5,000 L Gibco AGT media: 2 kg to 6,000 kg Select packaging based on workflow: Custom, or off-the-shelf for reduced lead time Closed system, ideal for aseptic processes and applications Solutions for disposable workflows Select QC testing, documentation: Standard QC testing: pH, osmolality, and sterility Documentation: Certificate of Analysis (COA), Material Safety Data Sheet (MSDS), import/export documentation Additional testing and documentation available upon request
Which custom media solution is best for you?

Gibco MediaExpressSM Services Description Regulatory Batch sizes Catalog modification or users formulation Non-cGMP Liquid: 1200 L Powder: 110 kg AGT media: 210 kg 2 weeks Liquid: 100 mL, 500 mL, 1 L bottles; 5 L, 10 L bags Powder/AGT media: 110 kg; range of plastic containers Check and record Formulation instructions lifetechnologies.com/custommedia Gibco Custom Cell Culture services Catalog modification or users formulation cGMP Liquid: 1010,000 L Powder: 18,000 kg AGT: 506,000 kg 1216 weeks Liquid: 100 mL, 500 mL, 1 L bottles; 1200 L range of bags Powder/AGT media: 110 kg; range of plastic containers; 55 gal HDPE drums Standard QA/QC release Certificate of Analysis (COA) lifetechnologies.com/custommedia Gibco PD-Direct Bioprocess Services Media development or optimization projects Non-cGMP Based on scope of work (SOW)
Lead time Packaging sizes
Variable Liquid: 100 mL, 500 mL, 1 L bottles; 5 L, 10 L bags Powder/AGT media: 110 kg; range of plastic containers Customizable Based on scope of work (SOW) Email: pd_direct@lifetech.com
Testing Documentation For more information
To get started, request a quote for custom media now. Customize a Gibco catalog media product at lifetechnologies.com/mediaconfigurator. Have us manufacture your own custom media formulation at lifetechnologies.com/custommedia.
31
Custom Novex multiplex assays for Luminex technology
New made-to-order assay design system: design and place an order or get a quote online
Our custom Novex multiplex immunoassay kits for the Luminex platform maximize flexibility in experimental design, permitting the quantitation of one or multiple targeted proteins in unique panels designed by you. For your convenience, each customized panel comes with reagents that are blended, optimized, tested, and designed for use with the Luminex 100/200 system, FLEXMAP 3D system, and the new MAGPIX system.
Custom Novex multiplex assays for Luminex technology
Choose specic targets of interest (polystyrene or magnetic)

Target Cytokine Chemokine Growth factor Signaling protein
Combined with appropriate Buffer Kit (all needed reagents) for ease of use
Species Human Mouse Rat Nonhuman primate
Each assay is premixed and QC tested for optimum top signal, low background, and linearity of dilution
Bead type Magnetic Polystyrene
Design and price your own Novex multiplex assay kit at bioinfo.invitrogen.com/luminex.
32
Custom biology services
Drug discovery demands custom research and custom solutions

From cell engineering to custom assay development, our scientists work closely with your team to understand your goals and tailor a solution that fits your unique project guidelines. When an off-the-shelf tool isnt the right option, our custom biology group offers unique and highly efficient solutions to help ensure you stay on track. Expect consistently short cycle times and high-quality results on time, every time, with proactive communication throughout the project.
Custom assays and services
Custom biochemical and cell-based assaysLet Life Technologies create a

custom biochemical or cell-based solution to suit your needs, using the broad array of technologies in the Life Technologies toolbox. Cell line generationOutsource your cell line development and let us craft a Jump-In parental cell line or final engineered cell line that meets your defined specifications. Antibody labelingChoose from europium or terbium donor chelate labels and obtain labeled antibodies for your TR-FRET assay development projects. Cell provisioningGet validated, high-throughput screeningready cells in large scale with our cryopreservation service. siRNA profilingIdentify and validate your target of interest using first-in-class siRNA collections with readouts to interrogate functional pathways, proliferation, gene expression analysis, and protein modifications. BacMam virus production assaysLet us create a BacMam virus from your gene of interest or develop a portable and robust BacMam-enabled assay. TaqMan protein assaysGet highly sensitive protein detection coupling the specificity of antibody affinity reagents with the sensitivity and fidelity of TaqMan PCR. Tell us your custom assay requirements at lifetechnologies.com/custom.
Custom biology services
Biochemical and cellular profiling services
Drug discovery scientists have trusted the SelectScreen Services team for thousands of screening and profiling projects on more than 500 targets. SelectScreen Services are executed with strict quality controls using a state-of-theart compound management system to drive down cycle times and meet expanding customer demand. Highly trained personnel and automated processes driven by integrated informatics systems help us ensure that reliable, high-quality data are delivered every time.
SelectScreen Kinase Profiling Serviceprofile with our expanding panel of SelectScreen Cell-Based Pathway Profiling Serviceunderstand potential
off-target effects earlier with our rapid, flexible cell-based profiling service against a panel of pathway-specific CellSensor cell lines. SelectScreen Cell-Based GPCR Profiling Servicechoose from a panel of functionally validated GeneBLAzer, second-messenger, and Tango betaarrestin recruitment cell-based assays. SelectScreen Cell-Based Nuclear Receptor Profiling Serviceprofile your compounds against the most comprehensive panel of target-specific GeneBLAzer nuclear hormone receptor assays for early safety and liability testing. Choose the right profiling service for your research at lifetechnologies.com/selectscreen. distinct protein kinases and get an average delivery time of just 6 days.
33
Gibco Cell Culture Basics
Instructional videos and companion handbook introduce the fundamentals of cell culture
The Gibco Cell Culture Basics videos and handbook provide critical training and troubleshooting for each essential cell culturing technique. Following these protocols will help your lab attain reproducible results every day. The handbook is an introduction to cell culture, covering the key topics listed below:
Cell culture laboratory safety Cell culture equipment Aseptic technique Aseptic technique checklist Biological contaminationtypes, cross-contamination,
Gibco Cell Culture Basics
Cell linesacquiring & selecting Adherent cell culture vs. suspension cell culture Cell culture environment (media, pH, CO2, and
temperature)
and use of antibiotics
Mammalian and insect cell morphology Cell culture protocols Guidelines for maintaining cultured cells Subculturing adherent cells Subculturing suspension cells Freezing cells Thawing frozen cells Counting cells in a hemocytometer Trypan blue exclusion Cell culture troubleshooting Cell culture products Transfection and selection
The video series includes all basic cell culture techniques through live demonstration. The video titles include:
Introduction to Cell Culture Sterile Technique Passaging Cells Freezing Cells Thawing Cells
Learn more about these videos and protocols at lifetechnologies.com/cellculturebasics.
Cell Line Database

The Cell Line Database is a Web-based tool that allows researchers to find more information about cell lines theyre already using, to find new cell lines, or to find compatible Life Technologies products and applications. The listing is alphanumeric and provides a rapid source of information about the type of cell, species, source type, and behavior of the cell in applications. The Cell Line Database can be found at lifetechnologies .com/celllinedatabase. Example search for HeLa cells
Technical ResourcesCell Line Database Cell Line Detail Cell Line Name: HeLa cells Tissues: Primary: Scientific application(s): Cervix No Transient Transfection Organism Type: Human Adherence Phenotype: Adherent
Products and applications for HeLa cells Gene Expression 7 Media/Cell Culture 15 Nucleic Acid Purification 1 Selective Antibiotics 1 Transfection Display 23 = Protocol exists within search results
34
Protocol Database
The Protocol Database is a Web-based tool that allows researchers to find more than 500 validated protocols for performing critical cell biology experiments. Protocols include culturing primary or stem cells, performing imaging and other cell analysis techniques, and protein purification, separation and identification techniques. The database allows the user to search for a specific protocol using keywords or to browse. The database can be found at lifetechnologies.com/protocolsdatabase.
Protocol Database
The Molecular Probes Handbook online

The online version of The Molecular Probes Handbook (lifetechnologies.com/handbook) is a comprehensive resource for fluorescence technology that is regularly updated. Like the printed version, the online Handbook contains detailed information about the thousands of Molecular Probes products available for fluorescence and imaging applications. The online Handbook has product pricing for your region as well as the added advantage of having critical product information at your fingertips. Find easy access to data images and referenced citations as well as product structures and spectra.
35
Fluorescence SpectraViewer
The Fluorescence SpectraViewer is an online tool that can be used to plot and compare the excitation and emission spectra of fluorescent dyes, facilitating the design of multicolor fluorescence experiments. As many as five fluorophores can be selected from an extensive list of dyes for simultaneous spectral analysis to determine the suitability of fluorophores for specific instrument configurations, and to evaluate spectral compatibility of different fluorophores. The SpectraViewer is available at lifetechnologies.com/spectraviewer.
Mobile applications and widgets

Life Technologies offers an assortment of handy applications and widgets for your everyday mobile needs. Download popular iPhone apps and widgets from Molecular Probes and other Life Technologies brands, including the DailyCalcs Science Calculator and the Alexa Fluor Selection Guide widget. Visit lifetechnologies.com/apps to get started. New apps and widgets are introduced on a regular basis, so check back often.
Virtual Cell Staining Tool

The Virtual Cell Staining Tool (lifetechnologies.com/ cellstainingtool) allows you to select different combinations of cellular structures and fluorophores to create your perfectly labeled fluorescent cell, which you can easily share with your colleagues using our email or print functionality.
36
Antibody and immunoassay selection guides

Product selection guide Immunoassay Selector Tool Labeling Chemistry Selector Tool Purpose Find the right Novex immunoassay product easily using our convenient selection tool. Simply search by keyword, then filter your results to find the right ELISA kit, Luminex platform assay, or antibody pair. http://igene.invitrogen.com/products/selector/immunoassays Quickly find the best Molecular Probes reactive dye or label for your experiment. Whether you want to label proteins and antibodies for immunofluorescence with our Alexa Fluor dyes, nucleic acids for in situ hybridization, or lipids for membrane studies, this tool will help you find the right conjugation product. http://igene.invitrogen.com/products/selector/dyes Primary Antibody Selector Tool Find all of the Life Technologies primary antibodies, including Molecular Probes flow cytometry antibodies, Novex primary antibodies for western blot analysis, and more. Simply search by target, gene symbol, or gene ID, then filter your results by application, reactivity, host, conjugate type, or antibody type. You can also search within your results to quickly find the right antibody. http://igene.invitrogen.com/isearch Secondary Antibody Selector Tool Finding the right secondary antibody is easier than ever. Whether you are looking for secondary antibodies, anti-dyes and anti-haptens, anti-epitope tags and anti-reporter genes, or isotype controls, Life Technologies has the largest selection available. Simply select the type of antibody you are looking for and filter by target, Ig class, reactivity, conjugate, or host. http://igene.invitrogen.com/antibody
Antibody and immunoassay selection guides
Customer and technical support

Comprehensive worldwide support
Whether you need help with a current order, placing a new order, or finding more information about product availability, please contact your local customer service team.
Technical support and training
If you have questions about product selection or use, assay or experimental design, data analysis, or troubleshooting, contact our team of technical support scientists or utilize our comprehensive portfolio of online product and application support tools.
LSIA 2011 Awards Most Responsive Customer Service & Most Knowledgeable Technical Support
How to reach us
To find your local customer service or technical support team, go to lifetechnologies.com/contactus. For product FAQs, protocols, training courses, and webinars, go to lifetechnologies.com/technicalresources. Follow us on Twitter: @LifeTechSupport @AppliedBio @dynabeads Like us on Facebook: facebook.com/appliedbiosystems facebook.com/lifetech.taqman facebook.com/gibcocellculture facebook.com/cellimaging facebook.com/immunologynews facebook.com/ambion facebook.com/invitrogen facebook.com/novexprotein @everydayprotein @gibco @invitrogen @molprobes @The_RNA_experts @lifeimmunology @LifetechEMEA
37
Gibco cell culture and cells

We have a 50-year heritage of bringing you dependable, consistent, and superior-quality cell culture products that provide scientists like you with the means and the confidence to elevate your research. From cutting-edge stem cell research tools like Essential 8 Medium, the CytoTune-iPS Sendai Reprogramming Kit, and our broad collection of human primary cells, to everyday essential media like DMEM and GlutaMAX Supplement, Gibco products help set the standard for cell culture in laboratories around the world.
Section Highlighted products
Primary and stem cells

Primary human keratinocytes, fibroblasts, corneal epithelial cells, umbilical vein endothelial cells, and more, page 56 Neural primary and stem cells, page 56, 71
Recover
Recovery Freezing Medium results in 25% more viable cells post-thaw and requires no additional supplementation, page 40
Grow
Gibco FreeStyle protein expression systems provide rapid, scalable, highyield transient expression in both 293 and CHO cells, page 42 Gibco CD Hybridoma Medium is a chemically defined, animal originfree medium optimized for the growth of hybridomas, page 42 Gibco Sf-900 III medium improves the consistency of your insect cell culture, page 47 Choose Geltrex, AlgiMatrix, or CellStart CTS 3D cell culture matrices to get more physiologically relevant culture conditions, page 49 Essential 8 Medium offers optimal feeder-free culture for all human PSC and iPSC lines with minimal lot-to-lot variability, page 60 StemPro NSC SFM is a serum-free medium for neural stem cells, page 52, 63 EpiLife medium for enhanced primary keratinocyte cell culture, page 57 Cell Therapy Systems (CTS) validated reagents for clinical research applications, look for the CTS label throughout this chapter
Mesenchymal stem cells, page 68 Hematopoietic stem cells, page 73
Everyday essentials
Classical Gibco media, cells, sera, and reagents for general cell culture in a wide variety of formats and sizes. Choose from standard media such as DMEM, RPMI, and MEM to more advanced formulations including GlutaMAX, OptiMEM, or Advanced reduced-serum media, page 41 Gibco provides FBS for both general and specialized cell culture, page 43 Discover a broad range of eukaryotic and bacterial selection antibiotics such as Geneticin, puromycin, and Zeocin, page 46 Gibco growth factors are manufactured for high activity, purity, and compatibility with Gibco media, page 47 B-27 Supplement, the most published serum substitute for neural cell culture, page 72 Neurobasal media, basal media formulated to sustain optimal culture of neurons when used with B-27, N-2, or G-5 supplements, page 71 KnockOut Serum Replacement, the standard supplement for growing pluripotent stem cells from multiple species, over 2,400 publications, page 6061
Instruments
38
The chart below highlights our most popular products and newest technologies in cell culture. Follow the Web links provided in this chapter for a complete listing of all our products and services. If you have any questions about which of our solutions are right for you, please contact Life Technologies technical support by phone, email, Facebook at facebook.com/gibcocellculture, or Twitter at twitter.com/gibco.
Passage
TrypLE Express, TrypLE Select, and StemPro Accutase are gentle and effective reagents for the dissociation of cell monolayers, page 52 TrypLE Select 10X is ideal for strongly adherent cells, page 52
Reprogram/engineer
The CytoTune-iPS Sendai Reprogramming Kit generates iPSCs from challenging cell types with this easy-to-use, nonintegrating technology, page 64 Episomal iPSC reprogramming vectors are integration-free and viral-free reprogramming vectors, page 64
Detect
Alkaline Phosphatase Live Stain is ideal for detecting pluripotency, page 66
Gibco liquid and powdered trypsin provide fast and convenient general-purpose cell dissociation, page 52
Countess Automated Cell Counter takes the subjectivity and tedium out of counting live and dead cells; fast, accurate, and convenient cell counting at your fingertips, page 53
39
Cell culture products

The Gibco brand of cell culture products has a 50-year heritage of dependability, consistency, and superior quality. Our goal is to provide scientists like you the means and the confidence to elevate research and healthcare now and into the future. We sincerely appreciate your trust, and are extremely proud to support your achievements thus farand the many yet to come. Explore our entire catalog of cell culture products at lifetechnologies.com/cellculture.
Recover
Efficient and effective cryopreservation is a critical step in cell culture maintenance. Recovered samples that have a large number of viable cells are the foundation of robust and healthy cultures, and allow you to begin experiments sooner and have more confidence in the results.
Grow
From the most basic formulations to the newest innovations, Gibco products are designed to provide the highest quality, consistency, and performance for your cell culture andtissue cultureneeds. These are media, reagents, cells, sera, and supplements from the leader in cell culture.
Passage
Gibco cell dissociation products are ideal for use with tissues and cell monolayers and come in a wide variety of formats to meet the diverse needs of researchers performing adherent cell culture. Automated cell counting improves the consistency of your cell culture passage to passage.
Recover
Recovery Freezing Medium Recovery Cell Culture Freezing Medium is a complete cryopreservation medium for mammalian cell cultures.
Recovery medium results in an average of 25% increase in cell viability in
Cell culture
Recovery medium is an optimized, fully supplemented formulation which avoids Recovery medium has proven performance on a broad range of mammalian Recovery medium contains DMEM, fetal bovine serum, calf serum, and 10%
DMSO. Recommended storage conditions: 5C to 20C Learn more at lifetechnologies.com/recovery. cell lines the messy mixing of DMSO
cryopreservation of both adherent and suspension cell lines
Grow
The following key cell culture interest areas are included in this section, with pages listed. Mammalian cell culture..............................4147 Insect cell culture........................................4748 Neurobiology media.....................................4849 Extracellular matrices.......................................49 Microbial culture................................................50 Cytogenetics.......................................................51
40
Mammalian cell culture

We offer products to support the growth of a variety of different mammalian cell lines, including ready-to-use options as well as powder and liquid formulations. Gibco cell culture media are time tested and cited in scientific literature more than any other brand in the world. Learn more about DMEM, RPMI Media 1640, MEM and other classical media at lifetechnologies.com/classicalmedia. Gibco Dulbeccos Modified Eagle Medium (DMEM) is designed to deliver superior quality, consistency, and control for your mammalian cell culture. We offer DMEM in a number of formulations. Gibco RPMI Media 1640 performs well in a wide range of applications for mammalian cells, including the culture of fresh human lymphocytes, fusion protocols, and growth of hybrid cells. We offer RPMI Media 1640 in a number of formulations. Gibco Minimal Essential Medium (MEM) is based on Eagles Media and is well suited for the growth of a broad spectrum of mammalian cells. We offer MEM in a number of formulations.
Classical media
GlutaMAX media
Gibco GlutaMAX medium is our standard, trusted cell culture medium that contains a stabilized dipeptide form of L-glutamine, L-alanyl-L-glutamine, which helps prevent degradation and ammonia build-up even during extended culture periods.
Minimizes toxic ammonia build-up Stable at room temperature Improves cell viability and growth
Extremely stable in aqueous solution, the L-alanyl-L-glutamine dipeptide will not degrade into ammonia in storage or incubation like L-glutamine. When cultured in medium that contains GlutaMAX-I supplement, cells gradually release aminopeptidases that hydrolyze the dipeptide, which in turn slowly releases L-alanine and L-glutamine into the culture media. The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. This is analogous to the fed-batch culture strategy in which L-glutamine is continuously added to the culture but maintained at a low concentration. The result is efficient metabolic processing and high growth yield without excess ammonia. We offer GlutaMAX-I supplement in DMEM, MEM, RPMI, Opti-MEM medium and others. You can also purchase the GlutaMAX-I dipeptide and substitute it directly for L-glutamine in your current cell culture media formulation. Learn more at lifetechnologies.com/glutamax.
Cell culture
Gibco Advanced reduced-serum media are enhanced basal media formulations of DMEM, DMEM/F-12, MEM, and RPMI 1640. Enriched with normal-serum constituents, these media require 5090% less FBS supplementation, deliver equivalent or superior cell growth compared to media supplemented with 10% FBS, and elicit no changes in morphology or function in many common cell lines. Learn more at lifetechnologies.com/advanced.
Advanced media
Gibco Opti-MEM reduced-serum medium is an improved minimal essential medium (MEM) that allows for a reduction of FBS supplementation by at least 50% with no change to growth rate or morphology. Opti-MEM medium is also recommended for use with cationic lipid transfection reagents, such as Lipofectamine reagent. Opti-MEM medium can be used with a variety of suspension and adherent mammalian cells, including Sp2, AE-1, CHO, BHK-21, HEK, and primary fibroblasts. Opti-MEM media are available in various formats for a range of cell culture applications, including hybridoma culture and transient and stable protein expression cultures. Learn more at lifetechnologies.com/optimem.
Opti-MEM media
41
Hybridoma media
Improve serum-free hybridoma culture with Gibco media
Choose from chemically defined, protein-free, and serum-free formulations Animal originfree products help minimize risk Custom products and packaging available Easy to scale up to large-volume cultures
Learn more about hybridoma media at lifetechnologies.com/hybridoma. that maximize performance
Product
CD Hybridoma Medium* (Cat. No. 11279023) Chemically defined medium Yes
CD Hybridoma AGT Medium* (Cat. No. 12372025) Chemically defined medium Yes
Hybridoma-SFM* (Cat. No. 12045084) Serum-free medium; low-protein (20 g/mL) No
PFHM-II* (Cat. No. 12040077) Protein-free medium Yes
Media type Animal originfree Optimized for
Human, mouse, rat hybridomas, myelomas. NS0, NS-1, and other steroid-dependent cells when used with 250X Cholesterol Lipid Concentrate (Cat. No. 12531018).
Human, mouse, rat hybridomas, myelomas
Applications
Growth and MAb production; can be used to express other proteins in engineered myeloma cell lines
* Drug Master File available. Dry granular format of CD Hybridoma Medium.
Cell culture
Protein expression media

Mammalian transient expression systems enable large-scale production of proteins and are ideal for the expression of human or other mammalian proteins because they generate recombinant proteins with closerto-native folding and post-translational modifications (i.e., glycosylation) than those expressed from E. coli, yeast, or insect cells. Gibco optimized mammalian protein expression media for CHO and 293 suspension cultures help you achieve rapid, scalable, high-yield protein production from transiently transfected cultures, and offer easy scale-up and purification with serum-free and animal originfree components. Learn more about the expression systems highlighted below at lifetechnologies.com/freestyle. Expi293 Expression Systemhigh-density, high-yield, scalable protein expression FreeStyle MAX 293 Expression Systemnew and improved FreeStyle MAX transfection reagent Each system supplies: FreeStyle 293-F cells FreeStyle 293 Expression Medium FreeStyle MAX Transfection Reagents pCMVSPORT -gal vector OptiPro SFM FreeStyle MAX CHO Expression systemlarge-scale protein expression in CHO cells Each system supplies: FreeStyle CHO-S cells FreeStyle CHO Expression Medium FreeStyle MAX Transfection Reagents pCMVSPORT -gal vector OptiPro SFM
Expi293-F cells Expi293 Expression Medium ExpiFectamine Transfection Kit pcDNA 3.4 vector Antibody-expressing positive control vector Opti-MEM I
FreeStyle 293 Expression Systemlow-density, large-scale protein expression Each system supplies: FreeStyle 293-F cells FreeStyle 293 Expression Medium 293Fectin Transfection Reagents pCMVSPORT -gal vector Opti-MEM I
Each system supplies:
42

Mammalian cell lines adapted to a variety of media formulations to meet your protein expression needs. Each lot is tested for cell growth and viability post-recovery from cryopreservation, and contains documented lineage from a low-passage Master Cell Bank. In addition, Master Seed Banks have been tested for contamination with bacteria, yeasts, mycoplasmas, and viruses and have been characterized by isozyme and karyotype analysis.
Source Mammalian Mammalian Mammalian Mammalian Cell type 293-F 293-F DG44 (Cells and Media Kit) CHO-S (Cells and Media Kit) Adapted media FreeStyle 293 and CD 293 CD 293 CD DG44 CD CHO Cat. Nos. R79007, 11625019 11631017 A1100001 A1369601 Suspension* cGMP-banked
* It is possible to achieve adherence in these cell lines by the addition of FBS or through the use of specially formulated media. Contact Technical Support for information.
FBS
Serum processed from bovine and other animals is a complex of over 10,000 components, including growth factors, proteins, lipids, minerals, and attachment factors, all of them useful for growth and maintenance of cells. Serum is a very cost-effective supplement added to basal media to make complete media for cell culture. The quality of serum is determined by its ability to grow healthy cells consistently over time and passages. Factors like sterility level, absence of microorganisms, level of endotoxin, ability to grow cells without downstream effects, and analysis of hormonal and biochemical profile of sera are important to scientists. Gibco sera are processed within a fully integrated operations system, controlled and supervised by Life Technologies personnel, right from the collection stage to processing to the cell culture hood. We process sera under stringent cGMP conditions at our ISO-certified, FDA-registered facilities. Most Gibco sera are labeled for in vitro diagnostics, compliant with the highest level of USP sterility testing, and triple-filtered at 0.1 m. Learn more at lifetechnologies.com/fbs. Our Gibco sera portfolio is classified into five product categories to help meet the needs of researchers based on cell types, specific assays, and downstream applications of cell culture.
Cell culture
An excellent value product for basic research with robust cell lines. Includes Qualified FBS products with endotoxin specification 50 EU/mL from USDA-approved regions, South America, Canada, and EU-approved regions.
Standard FBS
Performance FBS
Recommended for general cell culture with commonly used cell lines. Qualified FBS US-origin products with low endotoxin, specification 10 EU/mL.
Performance Plus FBS
For cell culture with a broad range of cell lines, especially for sensitive cell lines. Certified FBS US-origin products with the lowest endotoxin level, specification 5 EU/mL. These products undergo the highest level of testing, including biochemical and hormonal profiles, useful for analysis of downstream assays.
43

Choose the FBS thats right for your project and your budget. Find this interactive table online at lifetechnologies.com/fbs.
Standard Sera for cell culture with robust cell lines. Good value for basic research. FBS Qualified (USDA-approved, Canada, South America origin) IVD 50 EU/mL Standard testing Performance Low-endotoxin sera for general cell culture with common cell lines. FBS Qualified (US origin) IVD 10 EU/mL Standard testing Performance Plus Lowest endotoxin for a broad range of cell types, especially sensitive cell lines. FBS Certified (US origin) IVD 5 EU/mL Standard testing, plus analytical tests for hormone and biochemical profiles 16000044 FBS Certified (US origin) Heat-inactivated 1001,000 mL sizes 50 mL One Shot packs
Product Product use Endotoxin specification Quality/performance testing (gamma-irradiated upon request) Popular Cat. No. (500 mL) Convenient options
10437028 FBS Qualified (USDA-approved) Heat-inactivated 1001,000 mL sizes 50 mL One Shot packs
26140079 FBS Qualified (US origin) Heat-inactivated 1001,000 mL sizes 50 mL One Shot packs
Sera for cell culture with the least viral risk, sourced from BSE-free regions. Preferred for academic, industrial, and preclinical research requiring low BVDV.
Secure FBS
Cell culture
Exclusive screening for BVDV at collection Endotoxin 10 EU/mL for FBS and donor bovine sera Available as heat-inactivated FBS for ready-to-use convenience Pack sizes from 100 to 1,000 mL
Recommended Gibco sera: FBS, Qualified, Australia-origin (Cat. No. 10099141), and heat-inactivated version (Cat. No. 10100147). Find out more about secure FBS at lifetechnologies.com/securefbs.
Specialty sera
Sera specifically qualified for cell culture for specialty research and specific assays, including stem cell research, immunoassays, antibody production.
Embryonic stem cellqualified FBS Charcoal-stripped FBS Dialyzed FBS

Find out more at lifetechnologies.com/specialtysera.
These products include bovine, horse, newborn calf, goat, rabbit, porcine, and chicken sera, predominantly sourced from New Zealand, in pack sizes from 50 mL One Shot aliquots to 1,000 mL bottles. Find out more at lifetechnologies.com/otheranimalsera. Our animal serum products are offered in regular as well as heat-inactivated formats, in pack size options from the convenient 50 mL One Shot aliquots to 1,000 mL, up to 20 L packs for high-throughput cell culture. In addition, several custom-processed products are available for specific research needs, including gamma-irradiated and cell linescreened.
Other animal sera
We also offer our free, proprietary multiparametric sera lot matching tool, the iMATCH tool, to help you find the most consistent or highest-performing lot of serum available. Find the iMATCH tool at lifetechnologies.com/imatch.
iMATCH lot matching tool
44
Reagents
From beginning to end, Gibco cell culture reagents are manufactured according to the highest quality and service standards. From the most basic formulations to the newest innovations, Gibco customer-focused innovation helps ensure our products provide superior performance and consistencyfor results that you can count on every day. Find more information on the reagents in this section at lifetechnologies.com/culturereagents.
Balanced salt solutions can provide an environment that maintains the structural and physiological integrity of cells in vitro. All Gibco DPBS, PBS, HBSS, and EBSS balanced salt solutions are manufactured in stateof-the-art cGMP, ISO-certified facilities to offer the highest quality and consistency for reproducible results. Tests include osmolality, pH, stability, sterility, and endotoxin tests. All powdered balanced salt solutions are produced without sodium bicarbonate to increase stability.
Balanced salt solutions
HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic organic chemical buffering agent that is widely used to provide supplemental buffering to cell culture medium at pH 7.27.6, for example, when cell culture requires extended periods of manipulation outside of a CO2 incubator. The level of HEPES in cell culture media generally varies from 10 mM to 25 mM. In Gibco DMEM, it is at a concentration of 25 mM; in Gibco DMEM/F-12 it is at 15 mM.
HEPES buffer
Sodium bicarbonate (NaHCO3) is the most commonly used buffer for cell culture media. This is due to its nutritional benefits despite the reduced buffering capacity at physiological pH.
Sodium bicarbonate
Cell culture
Addition of amino acid supplements helps prolong the viability of cells in culture, stimulates growth, and reduces the biosynthetic burden on cultured cells. The nonessential amino acids in this solution are prepared in distilled water and provided at 100X concentration.
Amino acids
We produce membrane-filtered and endotoxin-screened distilled water in a number of packaging configurations and volumes for routine cell culture. We also offer Gibco Water for Injection (WFI) for Cell Culturea high-quality, cell culturegrade water that meets the United States Pharmacopeia monograph for water for injection packaged in bulk for commercial use elsewhere. Gibco Water for Injection (WFI) for Cell Culture is manufactured in ISO-certified, cGMP, and FDA-registered facilities.
Water
Media supplements are included in mammalian cell culture systems to help customize the growth conditions, improve the cell viability and growth, and keep cells healthier longer. We produce an extensive selection of media supplements that help ensure the reliability and consistency of your cell culture research. Find out more at lifetechnologies.com/mediasupplements.
Supplements
FoamAway Irradiated AOF is a ready-to-use antifoaming agent designed to offer high performance and convenience in combating foam in your cell culture system. It is prediluted, presterilized, and packaged to connect directly to your bioreactor with no further processing required. Find out more at lifetechnologies.com/foamaway.
Antifoaming agent
45

Antibiotics and antimycotics
For researchers, two types of cell culture contamination require careful monitoring: the contamination of cell cultures with microbiological organisms and the contamination of one cell line with another. Both forms of contamination are extremely prevalent and cannot be underestimated. The decision to use antibiotics and antimycotics to prevent contamination should be based on the individual researchers needs and experience. Life Technologies manufactures a wide array of antibiotics and antimycotics for contamination due to bacteria, fungi, or yeasts. While researchers may attempt to eliminate or control contaminations with antibiotics or antimycotics, it should be noted that these substances can be toxic to certain cell lines. Before using, it is recommended that you first perform a dose-response test to determine the level at which toxicity begins.
Gibco high-quality selection agentsprovide unique solutions for your research needs, such as dual selection and rapid stable cell line establishment. Choose the selection antibiotic that is best suited to your needs.
Selection antibiotics
Eukaryotic selection antibiotics. Find this interactive table online at lifetechnologies.com/selectionantibiotics.

Selection antibiotic Blasticidin Geneticin (G-418) Most common selection usage Eukaryotic and bacterial Eukaryotic Common working concentration 120 g/mL 100200 g/mL bacteria 200500 g/mL mammalian cells 200500 g/mL 25 g/mL 0.25 g/mL 50400 g/mL Available powder sizes 50 mg 1 g, 5 g, 10 g, 25 g Available liquid sizes 10 x 1 mL, 20 mL 20 mL, 100 mL
Cell culture
Hygromycin B Mycophenolic acid Puromycin Zeocin
Dual-selection experiments and eukaryotic Mammalian and bacterial Eukaryotic and bacterial Mammalian, insect, yeast, bacterial, and plant
500 mg
20 mL 10 x 1 mL, 20 mL 8 x 1.25 mL, 50 mL
Bacterial selection antibiotics. Find this interactive table online at lifetechnologies.com/selectionantibiotics.

Selection antibiotic Actinomycin D Ampicillin, sodium salt Blasticidin Carbenicillin, disodium salt Kanamycin sulfate Mycophenolic acid Neomycin sulfate Polymyxin B sulfate Puromycin Streptomycin sulfate Zeocin Most common selection usage Bacterial Bacterial Eukaryotic and bacterial Agrobacterium and E. coli Bacterial Mammalian and bacterial Bacterial Bacterial Eukaryotic and bacterial Bacterial Mammalian, insect, yeast, bacterial, and plant Common working concentration 1 g /mL 1025 g/mL 120 g/mL 100500 g/mL 100 g/mL 25 g/mL 50 g/mL 100 units/mL 0.25 g/mL 50100 g/mL 75400 g/mL Available powder sizes 5 mg, 10 mg 200 mg 50 mg 5g 5 g, 25 g 500 mg 100 g 25 MU Available liquid sizes 10 x 1 mL, 20 mL 100 mL 10 x 1 mL, 20 mL 100 g 8 x 1.25 mL, 50 mL
46
Growth factors
Gibco growth factors, chemokines, and cytokines are pure, high-quality proteins bioassayed with Gibco media, making them compatible with the cell culture media brand you use and trust. In addition, each protein is analyzed for purity along with structural homogeneity to help ensure a biologically active protein is produced.
On our website, youll find details on our broad selection of growth factors, cytokines and chemokines, including VEGF, EGF, FGFb, NGF, and SCF. Go to lifetechnologies.com/growthfactors to find:
Find more information online
Product groups narrowed alphabetically by name or by protein family or species Data demonstrating the purity, biological activity, and functionality of our recombinant proteins Information on our ISO 13485quality growth factors for clinical research
Insect cell culture

Insect cell culture is a common choice for heterologous protein expression. For large-scale production or basic research, insect cells are able to express large quantities of protein with complex posttranslational modifications. Gibco insect media have been formulated for maximum growth and protein yields. These media, in combination with Gibco preadapted cell lines, will provide a convenient system to save you time and effort.
Cell culture
Insect cell media selection guide. Find this interactive table online at lifetechnologies.com/insectcellculture.
Basic insect cell research Graces Insect Medium Serum free Protein free Source origin Lot-to-lot consistency Optimized to preventfoaming Compatible insectcells Supported cell density (Sf9cells/mL) Packaging Animal Low (FBS variability) Sf9 and Sf21 3 x 106 cells/mL 100 mL, 500 mL, 10 x 1L (powder) Sf9 and Sf21 10 x 106 cells/mL 500 mL, 1,000 mL, 5 L (bags), 10 L (bags), 20 L (bags) Serum-free Medium (medium hydrolysate concentration) Serum-free growth and protein expression Sf-900 II Highest consistency and protein expression Sf-900 III

Animal originfree High (low hydrolysate concentration)
Sf9 and Sf21
1014 x 106 cells/mL 500 mL, 1000 mL, 10 L (bags), 20 L (bags)
Drosophila S2 cells are used for heterologous protein expression in the Drosophila Expression System (DES). The S2 cell line was derived from a primary culture of late stage Drosophila melanogaster embryos (Schneider, 1972). This adaptable cell line grows without CO2, at room temperature, as a semi-adherent monolayer or suspension in Schneiders Drosophila Medium. Learn more at lifetechnologies.com/insectcellculture.
Drosophila S2 Cell Culture
High Five Cell Culture
High Five Cells (BTI-TN-5B1-4) are a clonal isolate, derived from the parental Trichoplusia ni cell line. They are optimized for the growth and maintenance of cells used with the Baculovirus expression vector system (BEVS) for adherent or suspension culture. Adapted to serum-free suspension culture in Gibco Express Five serum-free medium. Learn more about Express Five SFM (Cat. No. 10486025).
47

Insect cell lines adapted to a variety of media formulations to meet your protein expression needs. Each lot is tested for cell growth and viability post-recovery from cryopreservation, and contains documented lineage from a low-passage Master Cell Bank. In addition, Master Seed Banks have been tested for contamination with bacteria, yeasts, mycoplasmas, and viruses and have been characterized by isozyme and karyotype analysis.
Source Insect Insect Insect Insect Cell type Sf9 Sf21 High Five Drosophila S2 Adapted media Graces, Sf-900 II, and Sf-900 III Graces, Sf-900 II, and Sf-900 III Express Five SFM Schneiders Cat. Nos. B82501, 11496015, 12659017 B82101, 11497013, 12682019 B85502 R69007 Suspension Adherent
(Semi-adherent)
Neurobiology media
We offer an array of media for neural cell culture, including neural primary and neural stem cells, media, supplements, substrates, and growth factors. All of our products have been developed to work together for optimal performance as a complete cell culture system. Gibco media and supplements are the most widely used for neural cell culture. Gibco products are designed to provide the highest quality, consistency, and performancefor results you can count on.
Cell culture
Neurobasal media are a full line of basal media formulated to sustain optimal culture of neurons. Our Neurobasal media must be supplemented with either a serum-free supplement (such as B-27 Supplement or N-2 Supplement) or serum, and 0.5 mM L-glutamine or GlutaMAX-I Supplement.
Neurobasal media
Cited in many peer-reviewed publications Supports growth of neurons from the hippocampus, embryonic striatum, substantia nigra, septum,
cortex, neonatal dentate gyrus, and cerebellum when combined with B-27 Supplement
Choose the right Neurobasal media for your experimental needs from the list below:
Requirements For prenatal and fetal neurons For postnatal and adult brain neurons Recommended products Neurobasal Medium (1X) Minus Phenol Red (Cat. No. 12348017) Neurobasal Medium (1X) (Cat. No. 21103049) Neurobasal-A Medium (1X) Minus Phenol Red (Cat. No. 12349015) Neurobasal-A Medium (1X) (Cat. No. 10888022) Neurobasal-A Medium (1X) (Custom) (Cat. No. 0050128DJ) Neurobasal-A Medium (1X) Minus Phenol Red (Cat. No. 12349015) Neurobasal-A Medium (1X) (Cat. No. 10888022) Neurobasal-A Medium (1X) (Custom) (Cat. No. 0050128DJ) Neurobasal-A Medium (1X) (Custom) (Cat. No. 0050128DJ) Neurobasal Medium (1X) Minus Phenol Red (Cat. No. 12348017) Neurobasal-A Medium (1X) Minus Phenol Red (Cat. No. 12349015)
For tumor cell lines
No glucose/no sodium pyruvate No phenol red
Learn more at lifetechnologies.com/neurobasal.
48

Hibernate media
Hibernate media, when supplemented with B-27 Supplement and GlutaMAX-I Supplement, allow for the manipulation of neurons at ambient CO2 for at least 48 hours while retaining their viability. Hibernate media can also be used to preserve viable brain tissue for up to a month when stored at 4C and have also been shown to be suitable transport media to ship various tissues and biological specimens, including umbilical cord tissue. There are two types of Hibernate media, Hibernate-A and Hibernate-E, formulated to be used for postnatal and embryonic neurons, respectively. These two products have similar formulations except for a difference in osmolality: Hibernate-A has a higher range of osmolality than Hibernate-E. Learn more at lifetechnologies.com/hibernate.
Astrocyte medium
Gibco Astrocyte Medium (Cat. No. A1261301) has been specially formulated to support the growth of primary human and rat astrocytes while retaining their phenotypes. The medium has been internally validated and optimized to provide an optimal balance between cell growth and the maintenance of cell viability, normal phenotype, and function. If more rapid proliferation is desired, addition of an appropriate amount of epidermal growth factor (EGF) is recommended, although cells may exhibit morphological and phenotypic changes.
Extracellular matrices
Culturing cells on flat plastic surfaces results in artificial two-dimensional sheets of cells. In the in vivo state, human cells experience a three-dimensional environment, completely surrounded by other cells, membranes, fibrous layers, and adhesion proteins. Our Gibco extracellular matrices, scaffolds, and proteins produce a growth environment closer to that found in vivo, and this physiologically relevant environment allows you to get more realistic cell biology data. At lifetechnologies.com/3dcellculture, you can find information about:
Cell culture
Rat tail and bovine collagen I Natural mouse laminin to facilitate the attachment and expansion of ESCs, iPSCs, and NSCs Extracellular matrix (ECM) proteins, including fibronectin and vitronectin
Geltrex Matrix is a soluble form of reduced growth factor (RGF) basement membrane extract (BME) purified from murine Engelbreth-Holm-Swarm tumor.
Geltrex LDEV-free matrix products
Free of the lactate dehydrogenase-elevating virus (LDEV), making it ideal for all types of cell culture and
mouse in vivo research Tested for its ability to support endothelial tube formation from cryopreserved endothelial cells Available in standard and hESC-qualified formulations Learn more at lifetechnologies.com/geltrex.
The AlgiMatrix 3D Culture System is the first user-friendly, animal-free bioscaffold available for the development of higher-fidelity cell culture models that are more predictive of disease states and drug responses.
AlgiMatrix 3D cell culture system
Three-dimensional, porous alginate cell culture platform Chemically defined, animal originfree material Supports formation of 3D cellular aggregates that more closely reflect normal cell morphology and Easy visualization of cells Lyophilized product available in 6-well, 24-well, and 96-well plate formats
Learn more at lifetechnologies.com/algimatrix. behavior
49
Microbial culture
Microbial culture is a well-established research tool in molecular biology for the culture of bacteria and yeasts. Life Technologies offers a wide array of bacterial culture and yeast culture products, including growth media and reagents, to help you achieve your desired results. Learn more at lifetechnologies.com/microbialculture.
Liquid and powder growth media specifically formulated to culture bacteria for use in applications including cloning and protein expression.
Bacterial
Choose your type of bacterial growth medium.

LB broth and LB agar Terrific broth M9 minimal MagicMedia Medium ImMedia Medium Other media Most referenced bacterial medium for growth and maintenance of recombinant E. coli strains Nutritionally rich bacterial medium for higher-density growth and maintenance of recombinant E. coli strains Commonly used bacterial medium for cultivation and maintenance of E. coli strains Customized bacterial medium for growth of T7-regulated E. coli strains for autoinduction of protein expression Premixed, presterilized bacterial medium containing selection antibiotics for growth of E. coli strains Microbial media for outgrowth of transformed E. coli cells (S.O.C. medium), solid microbial media (Select Agar), and growth of hosts for replication of M13 vectors (2-YT Broth)
Learn more at lifetechnologies.com/bacterialmedia.
Cell culture
Yeast
Liquid and powder growth media specifically formulated to culture yeast for use in maintaining and propagating yeast strains.
Choose the appropriate yeast growth medium.

PichiaPink Media CSM medium YPD broth Yeast Nitrogen Base (YNB) Prepackaged media pouches for use with PichiaPink Yeast Expression System, containing media to grow P. pastoris yeast strains for transformation and selection Specialty medium for making agar plates to grow S. cerevisiae (MaV203) competent cells Commonly used yeast medium for maintenance and propagation of P. pastoris and S. cerevisae yeast strains Base medium for preparation of minimal and synthetic defined yeast media
Learn more at lifetechnologies.com/yeastmedia.
Algae
Gibco TAP Growth Medium (Cat. No. A1379801) optimized for Chlamydomonas reinhardtii, and Gibco BG-11 Growth Medium (Cat. No. A1379901) optimized for cyanobacteria. Learn more about algae protein expression systems at lifetechnologies.com/algaekit.
50
Cytogenetics
Gibco cytogenetics products are specifically formulated to help deliver clear, reproducible results and are application-tested by an independent laboratory. Each medium has been designed and optimized for the analysis of amniotic fluid cells, chorionic villus samples, bone marrow cells, or peripheral blood lymphocytes.
Optimized and prequalified for cytogenetics Provide high mitotic index Deliver excellent chromosomal morphology Offer clear, reproducible results that are easy to analyze and interpret Product use: IVD
Learn more at lifetechnologies.com/cytogenetics.
Gibco AmnioMAX-C100 and AmnioMAX II complete media are designed to increase cell attachment and growth rates and provide high metaphase yields. Learn more at lifetechnologies.com/amniomax.
AmnioMAX media
MarrowMAX Bone Marrow Medium
A bone marrow aspirate provides unique and valuable research data, but cell numbers are generally very lowtherefore, the medium you select could not be any more critical. MarrowMAX Bone Marrow Medium contains a novel stromal cellconditioned medium for optimal growth and minimizes the need to supplement media, helping to save both time and money. MarrowMAX medium far outperforms commercially available formulations supplemented with giant cell tumor (GCT)conditioned media, offering consistent lot-to-lot performance with a higher mitotic index and superior chromosome morphology. Learn more at lifetechnologies.com/marrowmax.
Cell culture
PB-MAX Karyotyping Medium
PB-MAX Karyotyping Medium is a fully supplemented, RPMI 1640based medium optimized for the karyotype analysis of peripheral blood lymphocytes. Ready to use, it is proven for performance and reliability. Learn more at lifetechnologies.com/pbmax.
The KaryoMAX line of products is designed to reduce culture time and days to harvest. Our line of KaryoMAX reagents includes KaryoMAX Colcemid, KaryoMAX Giemsa Stain, and KaryoMAX potassium chloride solution. Learn more at lifetechnologies.com/karyomax.
KaryoMAX products
51
Passage
Gibco cell dissociation products are ideal for use with tissues and cell monolayers. Gibco trypsin and TrypLE cell dissociation reagents come in a wide variety of formats to help meet the diverse needs of researchers performing adherent cell culture.
Cell dissociation reagent selection guide. Find this interactive table online at lifetechnologies.com/celldissociation.
Fast, general purpose Trypsin Source origin Animal origin (porcine) Frozen (5 to 20C) Gentle and convenient for research use TrypLE Express (1X) Animal originfree Gentle and convenient for bioproduction/ industrial applications TrypLE Select (1X) Animal originfree (dedicated AOF machinery) Ready to use (room temperature stable) Gentle and fast for strongly adherent cells TrypLE Select (10X) Animal originfree (dedicated AOF machinery) Ready to use (room temperature stable) Specialized for neural and pluripotent stem cell dissociation StemPro Accutase Serum-free, animal origin (marine invertebrate) Refrigerated (2 to 8C)
Storage temperature Higher cell viability Inactivation method Available packaging
Ready to use (room temperature stable)
Requires trypsin inhibitors/neutralizers 100 mL, 500 mL, 100 g powder Inhibition by dilution (no inhibitors needed) 100 mL, 500 mL, 5 L bags
Inhibition by dilution (no inhibitors needed) 100 mL, 500 mL
Inhibition by dilution (no inhibitors needed) 100 mL, 500 mL 100 mL
Inhibition by dilution (no inhibitors needed)
Cell culture
52
Automated cell counting

The Countess Automated Cell Counter takes the subjectivity and tedium out of one of the fundamental steps of cell culture counting cells and assessing viability. The Countess Automated Cell Counter helps to eliminate subjectivity and minimize human error for comprehensive, reproducible data. The Countess Automated Cell Counter is:
Accurateeliminate the subjectivity of manual cell counting; no guessing, no user-to-user variability Fastcounts live and dead cells and measures viability and average cell size, typically in 30 seconds with just 10 L of sample Convenientno setup, cleaning, or service required
The Countess Automated Cell Counter is easy to use. Simply pipet the sample into the counting slide, insert the slide into the Countess Automated Cell Counter, then press Count cells; results are displayed in 30 seconds. See the Countess Automated Cell Counter in action at lifetechnologies.com/countess.
Easy workflow and accurate results
Cell culture
The Countess Automated Cell Counter workflow. (A) Pipet the sample into the counting slide. (B) Insert the slide into the Countess Automated Cell Counter. (C) Press Count cells. (D) Results are displayed in 30 seconds.
53
Human primary cells

Our portfolio of Gibco primary cell cultures and complete primary cell culture systems (including cells, media and supplements, matrices, cryopreservation medium, and analysis tools) give you the potential to generate more physiologically relevant data. Gibco complete cell culture systems are:
Optimized for each cell type Easy to use, offering single-shot supplementation Available in custom solutions for unique research needs
We maintain stocks of the following primary cells:
Keratinocytes Fibroblasts Melanocytes Corneal epithelial cells Microvascular entothelial cells Smooth muscle cells Large vessel endothelial cells (aortic, pulmonary, and umbilical) Hepatocytes Astrocytes Skeletal myoblasts Mammary epithelial cells
Browse our entire selection human primary cells at lifetechnologies.com/primarycells.
Primary cells
Life Technologies welcomes requests for custom preparations of cell culture products and contract research. Please contact your account manager or technical sales specialist, and we will work with you to develop a solution that meets your research and budgetary needs. Custom cell culture products and services
Custom primary cells and media
Custom cell isolations and configurations Custom medium and supplement formulations Cell pellets suitable for RNA isolation and other purposes Additional cell characterization and virus testing
Research applications for primary cells

Basic structure/function studies Dermal modeling Gene regulation Signal transduction Cell co-culturing Cancer biology Angiogenesis Melanoma Normal controls Drug discovery/cosmetics/beauty and personal care studies Acne HTS/HCA screening Pigmentation Secondary and tertiary screenings Toxicology screening In vitro alternatives to animal testing Corrosivity Cosmetics and topicals Household product irritancy Safety assessment testing services and products Regenerative medicine/ therapy Burn therapy Chronic skin ulcers Cosmetic (wrinkles, scars, hair growth) Wound healing
54

Products for engineering and analyzing primary cells
Life Technologies offers a host of products that enable further engineering of primary cells, as well as analysis of activity and function within primary cell models. These include detection technologies such as CellLight and Premo reagents based on BacMam technology, which enable efficiently delivery and gene expression in mammalian cells, even for the most difficult-totransfect primary cells. Additionally, our instrument platforms can simplify everything from counting, transfecting, and visualizing primary cells. These platforms include:
Neon Transfection System (lifetechnologies.com/neon) Tali Image-Based Cytometer (lifetechnologies.com/tali) FLoid Cell Imaging Station (lifetechnologies.com/floid) Attune Acoustic Focusing Cytometer (lifetechnologies.com/attune)
Cultured in EpiLife Medium + HKGS
Cultured in Medium 154 + HKGS
Primary cells
HEKn, day 1
HEKn, day 3
HEKn, day 5
Comparison of HEKn (Cat. No. C-001-5C), secondary culture, grown in either EpiLife Medium or Medium 154.
HCEC Growth Rates
B
Population doublings/day
1.2 1.0
3 culture 5 culture 7 culture
0.5
KSFM Growth Medium
dKSFM + CM
Fluorescent multiplex imaging of microfilaments in human dermal fibroblasts. (A) Phalloidin Alexa Fluor 488 (green); Hoechst 33342 (blue). (B) Antialpha/ tubulin/goat anti-rabbit Alexa Fluor 555 (red); phalloidin Alexa Fluor 488 (green); Hoechst 33342 (blue).
HCEC were thawed and seeded according to product instructions in Keratinocyte SFM (KSFM) or Defined Keratinocyte SFM (dKSFM). Cells were passaged at 90% confluence, and population doublings per culture were calculated. Bars show the mean growth in triplicate T-25 flasks, with standard deviation.
55

Life Technologies human primary cell systems quick reference guide
Anatomical location of derivation Skin Cells Human Epidermal Keratinocyte, neonatal (animal productfree) Human Epidermal Keratinocyte, adult (animal productfree) Human Epidermal Keratinocyte, neonatal Human Epidermal Keratinocyte, adult Human Epidermal Keratinocytes, pooled Human Melanocyte-neonatal, lightly pigmented Human Melanocyte-neonatal, moderately pigmented Human Melanocyte-neonatal, darkly pigmented Human Melanocyte-adult, lightly pigmented Human Dermal Fibroblasts, neonatal Human Dermal Fibroblasts, adult Human Microvascular Endothelial Cell, neonatal dermis Human Microvascular Endothelial Cell, adult dermis Heart Human Aortic Endothelial Cell Human Pulmonary Artery Endothelial Cell Human Aortic Smooth Muscle Cell Human Coronary Artery Smooth Muscle Cell Human Pulmonary Artery Smooth Muscle Cell Cornea Quadriceps Umbilical cord Human Corneal Epithelial Cell Human Skeletal Myoblasts, small size Human Skeletal Myoblasts, large size Human Umbilical Vein Endothelial Cell Human Umbilical Vein Endothelial Cell (pooled, 500,000 cells) Human Umbilical Vein Endothelial Cell (pooled, 1 million cells) Breast Human Mammary Epithelial Cell HUVEC HUVEC HUVEC HMEC Acronym HEKn-APF HEKa-APF HEKn HEKa HEKp HEMn-LP HEMn-MP HEMn-DP HEMa-LP HDFn HDFa HMVECnd HMVECad HAEC HPAEC HASMC HCASMC HPASMC HCEC HSkM Cryopreserved product Cat. No. C0205C C0215C C0015C C0055C A13401 C0025C C1025C C2025C C0245C C0045C C0135C C0105C C0115C C0065C C0085C C0075C C0175C C0095C C0185C A12555 A11440 C0035C C0155C C01510C A10565
Primary cells
Brain
Gibco Human Astrocytes Gibco Human Astrocyte Kit
NA
N7805100 N7805200
* Also available in Calcium Free and Calcium Free/Phenol Red Free Kits: EpiLife CF (Cat. No. MEPICF500) and CF/PRF (Cat. No. MEPICFPRF500), Medium 154 CF (Cat. No. M154CF500) and CF/PRF Kits (Cat. No. M154CFPRF500) Requires plating with Coating Matrix (Cat. No. R011K) for efficient cell attachment Also available as Calcium Free Kit: Medium 254CF Kit (Cat. No. M254CF500) Animal productfree supplements ** Also available as a Phenol Red Free: Medium 200PRF (Cat. No. M200PRF500) Requires plating with Geltrex matrix for efficient cell attachment.
56
Growth medium EpiLife* EpiLife*
Cat. No. MEPI500CA MEPI500CA
Growth supplement S7 EpiLife Defined Growth Supplement Human Keratinocyte Growth Supplement Human Keratinocyte Growth Supplement Kit
Acronym S7 EDGS HKGS HKGS kit HKGS HKGS kit HMGS
Cat. No. S0175 S0125 S0015 S001K S0015 S001K S0025
Medium 154* Medium 254
M154500 M254500
Human Keratinocyte Growth Supplement Human Keratinocyte Growth Supplement Kit Human Melanocyte Growth Supplement
Medium 254 Medium 106 Medium 131 (w/ Attachment Factor) Medium 200** Medium 231
M254500 M106500 M131500 M200500 M231500
Human Melanocyte Growth Supplement-2 Low Serum Growth Supplement Low Serum Growth Supplement Kit Microvascular Growth Supplement Large Vessel Endothelial Supplement Smooth Muscle Cell Growth Supplement Smooth Muscle Cell Differentiation Supplement
HMGS-2 LSGS LSGS Kit MVGS LVES SMGS SMDS
S0165 S00310 S003K S00525 A1460801 S00725 S0085
Primary cells
KSFM dKSFM DMEM, Low Glucose, Pyruvate Medium 200**
17005042 10744019 11885084 M200500
Note: Supplements included with media 2% Horse Serum Large Vessel Endothelial Supplement LVES 16050130 A1460801
Medium 171 HuMEC Ready HuMEC Basal Serum Free Medium Gibco Astroctye Medium
M171500 12752010 12753018 A1261301
Mammary Epitehlial Growth Supplement Note: Supplements included with media HuMEC Supplement Kit NA NOTE: Included in kit
MEGS NA
S0155 12755013
57

Life Technologies human primary cell systems quick reference guide
Anatomical location of derivation Liver Cells HEP10, Pooled Human Cryopreserved Hepatocytes Human Suspension Hepatocytes, Metabolism Qualified, Male Human Suspension Hepatocytes, Metabolism Qualified, Female Human Suspension Hepatocytes, Metabolism Qualified, Male, 912 million Human Suspension Hepatocytes, Metabolism Qualified, Female, 912 million Human Suspension Hepatocytes, Polymorphic Donors Human Suspension Hepatocytes, Polymorphic Donors, 912 million Human Suspension Hepatocytes, Transporter Qualified, 48 million Human Suspension Hepatocytes, Transporter Qualified, 912 million Human Plateable Hepatocytes, Induction Qualified Human Plateable Hepatocytes, Metabolism Qualified Human Plateable Heptocytes, Transporter Qualified Human Plateable Heptocytes, Uptake Qualified Cryopreserved product Cat. No. HMCS10 HMCS1S HMCS2S HMCS1L HMCS2L HMCSPS HMCSPL HMCSTS HMCSTL HMCPIS HMCPMS HMCPTS HMCPUS
Primary cells
58
Thawing reagents Cryopreserved Hepatocyte Recovery Medium (CHRM)
Cat. No. CM7000
Plating reagents Williams Medium E (1X, no phenol red) Hepatocyte Plating Supplement Pack (serum-containing) Collagen I, Coated Plates (6-well) or Collagen I, Coated Plates (24-well) or Collagen I, Coated Plates (96-well) + Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix
Cat. No. A1217601 CM3000 A1142801 A1142802 A1142803 A1413202
Maintenance reagents Williams Medium E (1X, no phenol red) Hepatocyte Maintenance Supplement Pack (serum-free) Collagen I, Coated Plates (6-well) or Collagen I, Coated Plates (24-well) or Collagen I, Coated Plates (96-well) + Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix
Cat. No. A1217601 CM4000 A1142801 A1142802 A1142803 A1413202
Primary cells
59
Pluripotent stem cells (PSCs)

Culture
Life Technologies offers an extensive range of Gibco media, supplements, and reagents to help meet all of your stem cell culture needs. Culture and expand stem cells in cGMP-manufactured serum-free, feeder-free, xeno-free cell culture systems. Manufactured with superior quality standards, our products offer the consistency and accuracy you need to get the most from your experiments. Find out about all of the options for PSC culture at lifetechnologies.com/culturepsc.
PSC culture medium selection guide.

The standard for serum-free, feeder-based culture of ESCs and iPSCs from multiple species Feeder-free culture with flexibility to optimize bFGF concentration and maintain differentiation capacity of PSCs StemPro hESC SFM A1000701 RUO Human; feeder-free culture Optimal feeder-free culture of all human ESC and iPSC lines with minimal lot-tolot variability Fully documented xeno-free PSC culture reducing the burden in qualifying reagents, facilitating transition from bench to clinic
Product Cat. No. Product use Recommended applications
KnockOut Serum Replacement 10828028, 10828010 Class II Medical Device Multiple species (including human and mouse); feeder culture; reprogramming on feeders; differentiation of PSCs Medium Compact colonies with smooth edges; high nuclear-to-cytoplasmic ratio
Essential 8 Medium KnockOut CTS XenoFree ESC/iPSC Kit A14666SA RUO Human; feeder-free, xeno-free culture; feeder-free reprogramming Low Compact colonies with smooth edges; high nuclear-tocytoplasmic ratio 1 x 10 mL Essential 8 Supplement; 1 x 500 mL DMEM/F-12 with GlutaMax-I A1448801, A1448701 RUO Human; feeder-based and feeder-free xeno-free culture for cell therapy research Medium Compact colonies with smooth edges; some colonies will have undefined borders with spiky edges 1 x 100 mL KnockOut SR XenoFree CTS 1 x 500 mL KnockOut DMEM CTS 1 x 5 mL KnockOut SR GF Cocktail CTS
Lot-to-lot variability Cell morphology
Medium Compact colonies with smooth edges; some colonies will have undefined borders with spiky edges 1 x 10 mL StemPro Supplement; 1 x 500 mL DMEM/F-12 with GlutaMax-I; 1 x 40 mL 25% BSA bFGF (Cat. No. PHG0264) 2-mercaptoethanol (Cat. No. 21985023)
Components/unit
1 x 100 mL or 500 mL
Additional components required for complete medium
DMEM/F-12 (1X) with GlutaMAX-I (Cat. No. 10565018) NEAA (Cat. No. 11140050) bFGF (Cat. No. PHG0264) 2-mercaptoethanol (Cat. No. 21985023)
For cost-effective option, choose the KnockOut ESC/iPSC Media Kit (Cat. No. A1412901)
No additional compo- bFGF CTS (Cat. No. CTP0263) nents required 2-mercaptoethanol (Cat. No. 21985023)
Stability of complete Stable at 4C for 2 weeks medium Specifically designed for cell therapy research Available support resources No
Stable at 4C for 2 weeks No
Stable at 4C for 2 weeks No
Stable at 4C for 2 weeks Yes
Protocols for culturing PSCs, publications, hands-on training, product finder
Protocols for culturing PSCs
Protocols for Protocols for culturing PSCs, culturing PSCs, FAQs, FAQs, product documentation hands-on training
60

KnockOut Serum Replacement (KSR) Cat. No. 10828010, 10828028
Well-referenced, proven standard
KnockOut Serum Replacement and basal media deliver confidence, with over 2,400 peer-reviewed publications.
Flexible feeder-free culture
StemPro hESC SFM Cat. No. A1000701
StemPro hESC SFM optimizes bFGF concentration and maintains the differentiation capability of PSCs.
Essential 8 Medium Cat. No. A14666SA

Feeder-free culture for all Essential 8 Medium contains only the necessary components for optimal PSC culture for every step of your research.
KnockOut CTS XenoFree ESC/iPSC Kit Cat. No. A1448801, A1448701

Traceability for cell therapy research The KnockOut CTS XenoFree ESC/iPSC Kit helps reduce the burden in qualifying reagents, facilitating transition from bench to clinic.
KnockOut ESC/iPSC Media Kit Cat. No. A1412901, A1413001

Bundled reagents for convenience
A cost-effective and convenient way to purchase the reagents required for feeder-based culture of human and nonhuman embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs). This kit includes DMEM/F-12 with GlutaMAX-I, KnockOut Serum Replacement, and FGF-basic.
KnockOut basal media

KnockOut DMEM Cat. No. 10829018 Optimized for growth of undifferentiated ESC and iPSCs
KnockOut DMEM is a chemically defined, reduced-osmolality basal medium designed for use with KnockOut Serum Replacement.
KnockOut DMEM/F-12 Cat. No. 12660012 Accelerates growth of ESCs and iPSCs
KnockOut DMEM/F-12 is a chemically defined, low-osmolality basal medium for use with KnockOut Serum Replacement.
Gibco growth factors are designed to provide the activity, stability, and validation required for your stem cell research.
Basic Fibroblast Growth Factor (bFGF) Cat. No. PHG0264
High biological activity and high puritymore results with less protein Proven compatibilityGibco proteins are bioassayed with Gibco media Find bFGF and other growth factors at lifetechnologies.com/growthfactors
61
Matrix products and attachment factors

We provide extracellular matrices (ECMs) that are designed to minimize adaptation time, maximize cell performance, and help meet regulatory requirements. Whether you are culturing your cells on feeder layers or require feeder-free xeno-free conditions for more clinical applications, Gibco products provide you with all the choices to culture your pluripotent stem cells (PSC) in the right environment. Use the guide below to help you choose the right product for your cells.
ECM and attachment factor selection guide.

Most commonly used basement membrane matrix for ESC and iPSC cultures Geltrex LDEV free hESC Qualified A1413301 RUO Feeder and feeder-free culture StemPro hESC SFM Murine No Frozen (20 to 80 C) 1 hr incubation at 37C 2 weeks at 28C 1 mL or 5 mL 16 x 6 well plates from 1 mL vial diluted 1:100 Optimized for use with Essential 8 Medium Vitronectin (VTN-N) A14701SA RUO Feeder-free and xeno-free culture Essential 8 Medium Human (recombinant) Yes Frozen (20 to 80 C) 1 hr incubation at room temp 1 week at 28C 1 mL 16 x 6-well plates from 1 mL Optimal for clinical research applications that require human origin only materials CELLstart CTS A1014201 RUO Feeder-free, xeno-free culture KnockOut SR XenoFree CTS supplemented medium Blend of human native proteins Yes Refrigerate (2 to 8C) 12 hr incubation at 37C 17 days at 28C (tightly sealed) 2 mL 22 x 6-well plates, when diluted 1:50
Product Cat. No. Product Recommended applications Recommended medium pairing Source origin Defined Storage temperature Plate preparation time
Shelf life of prepared plate Unit size Number of plates coated/unit
Note: Attachment Factor (0.1% Gelatin, Cat. No. S006100) may be used for feeder-layer attachment.
62
Passaging PSC cultures

We offer options for both enzymatic and mechanical passaging of PSC cultures. Which method you choose depends on the condition of the culture at the time you want to passage and what will be done with the cells after passaging. Use the guide below to help you select the right method.
Passaging method selection guide.

Recommended for clump passaging in feeder and feeder-free cultures Product Dispase Standard reagent for clump passaging in feeder and feeder-free cultures Collagenase IV Quick and gentle dissociation, recommended for singlecell passaging Recommended for clump passaging with feeder-free Essential 8 Medium UltraPure 0.5 M EDTA Optimal for clinical research applications that require human origin only materials TrypLE Select CTS Quick and easy manual passaging tool for feeder and feeder-free cultures StemPro EZPassage Disposable Stem Cell Passaging Tool 23181010 Mechanical passaging in clumps NA
StemPro Accutase
Cat. No. Recommended applications Source origin Form Storage temperature Shelf life of liquid format Inactivation method
17105041 Passaging in clumps Bacillus polymyxa Lyophilized powder 28C 2 weeks Inhibition by dilution (no inhibitors needed) 5g 2 mg/mL
17104019 Passaging in clumps Clostridium histolyticum Lyophilized powder 28C 2 weeks Inhibition by dilution (no inhibitors needed) 1g 1mg/mL for feeder-dependent, 10 mg/mL for feeder-free 500 x 60 mm plates at 1 mg/mL, 50 x 60 mm plates at 10 mg/mL
A1110501 Dissociation to single cells Marine invertebrate Ready-to-use liquid 28C 2 years Inhibition by dilution (no inhibitors needed)
15575020 Passaging in clumps Chemical, AOF Liquid, dilute 1,000x Room temperature 2 years Inhibition by dilution (no inhibitors needed) 4 x 100 mL 0.5 mM
A1285901 Single-cell passaging Recombinant enzyme (AOF) Ready-to-use liquid Room temperature 2 years Inhibition by dilution (no inhibitors needed) 100 mL 1X solution
Individually wrapped Room temperature NA NA
Unit size Optimized concentration for use
100 mL 1X solution
Box of 10 units NA
Number of plates passaged/unit
1,250 x 60 mm plates
50 x 60 mm plates
50,000 x 60 mm plates (100 mL at 1,000X dilution)
50 x 60 mm plates
10 plates
63
Tools for reprogramming

Selecting the right reprogramming tool to generate induced pluripotent stem cells (iPSCs) can leave you with questions. And no one has more answers than Life Technologies. From traditional methods (lentivirus) to nonintegrating technologies (Sendai virus and episomal vectors), Invitrogen products provide the breadth and depth you depend on. That means more solutions to help meet your safety, efficiency, and budgetary needs. Learn more at lifetechnologies.com/cellreprogram.
Lentiviral iPSC reprogramming particles

Well-referenced, flexible technology Choose from eight different high-titer lentiviral iPSC reprogramming particles for your custom iPSC experiments. Learn more at lifetechnologies.com/lenti.
Episomal reprogramming vectors Cat. No. A14703

Safety with confidence Create reproducible iPSCs from normal and diseased cell types with Invitrogen episomal vectors.
CytoTune-iPS Sendai Reprogramming Kit

High efficiency for difficult cell types
Reprogram challenging cell types with the easy-to-use, nonintegrating CytoTune-iPS Sendai Reprogramming Kit. Learn more at lifetechnologies.com/cytotune.
Reprogramming tool selection guide.

Well-referenced technology for iPSC generation from fibroblasts Product Cat. No. Lentiviral iPSC reprogramming particles A1357101, A1357201, A1357301, A1357401, A1357501, A1357601, A1357701, A1357801 0.0010.01% No No One application, selection and QC of putative colonies required No hOct4-GFP and CMV-GFP reporters available for transduction and expression verification Standard protocol for fibroblast reprogramming Proven technology for viral-free iPSC generation from normal and diseased cell types Episomal iPSC Reprogramming Vectors A14703 Efficient generation of integration-free iPSCs from multiple somatic cell types CytoTune-iPS Sendai Reprogramming Kit A1378001, A1378002
Reprogramming efficiency Genomic integrationfree Virus-free reprogramming Hands-on time Blood cell reprogramming Convenience
0.0020.08% Yes Yes One application, minimal QC of putative colonies required No Premixed, 3-plasmid kit; requires Neon Transfection System (or similar device) Protocols for fibroblast reprogramming
0.011% Yes No One application, minimal QC of putative colonies required Yes Kit contains all 4 genes required; cannot be purchased separately Protocols for fibroblast, CD34+, PBMC reprogramming. FAQs, webinar, publications
Available support resources
64
Other reprogramming tools

TaqMan iPSC Sendai Detection Kit Cat. No. A13640
These TaqMan probes are used for detecting the presence and determining the levels of Sendai virus and exogenous transcription factors (OCT 3/4, SOX2, KLF4, and c-Myc) delivered by the Sendai virus from the CytoTune-iPS Sendai Reprogramming Kit. The primers in the TaqMan iPSC Sendai Detection Kit will not detect the corresponding endogenous factors.
Primary cells for reprogramming

Primary cells are cells taken directly from living tissue and established for growth in vitro. These cells have undergone very few population doublings and are therefore more representative of the main functional component of the tissue from which they are derived Find all of our primary cells at lifetechnologies.com/primarycells.
Fibroblasts
Primary human dermal fibroblasts from either adult or neonatal samples.
Keratinocytes CD34+ cells
Primary human keratinocyte cells from either adult or neonatal samples.
Pooled human hematopoietic progenitor cells derived from the umbilical cord blood of mixed donors.
Shockingly simple transfection in stem cells
Neon Transfection System Cat. No. MPK5000
A next-generation electroporation technology for highly efficient delivery (~80%) of nucleic acids such as episomal vectors into somatic cells. Learn more at lifetechnologies.com/neon.
65
Detection
Validation is critical for your induced pluripotent stem cell (iPSC) research. Whether you want to quickly confirm that cells are pluripotent or need flexibility in antibody and dye choice, we have Molecular Probes labeling and detection assays and imaging stations to help you get the confirmation you need. And, all of our labeling and detection technologies are backed by our extensive internal and external support resources, including the Molecular Probes online community and the trusted Molecular Probes Handbook. Learn more at lifetechnologies.com/detectpsc.
Alkaline Phosphatase (AP) Live Stain shows robust staining in pluripotent stem cells. H9 human ESC grown on MEF feeders show specific staining of the pluripotent stem cell colonies.
Alkaline Phosphatase Live Stain Cat. No. A14353

Quick check for pluripotency Rapidly detect pluripotency in expanding ESCs and iPSCs without compromising cell integrity with Alkaline Phosphatase Live Stain. Learn more at lifetechnologies.com/stemcellanalysis.
Antibodies
Flexibility and choice Mix and match your choice of primary antibody and Molecular Probes fluorophore for maximum flexibility in pluripotent stem cell imaging.
H9 hESCs labeled with pluripotent surface antibodies: SSEA4Alexa Fluor 488 (green), TRA-1-81Alexa Fluor 594 (red), TRA-1-60Alexa Fluor 647 (purple).
Learn more at lifetechnologies.com/antibodies.
Cell imaging right at your benchtop
FLoid Cell Imaging Station Cat. No. 4471136
Confidently detect and validate your ESC and iPSC colonies with Live Alkaline Phosphatase and cell-surface antibodies on the affordable, user-friendly FLoid Cell Imaging Station. Learn more at lifetechnologies.com/floid.
66

Detection method selection guide.
Allows easy identification of pluripotency Allows easy identification of pluripotency without compromising cell integrity Alkaline Phosphatase Live Stain A14353 Medium (stains stem and progenitor cells) No Stains PSCs typically in 20 min Green/fluorescein (FITC) Specific and flexible identification of PSCs; antibodies can be combined with different fluorophores for co-staining Primary antibodies lifetechnologies.com/antibodies High (stain human ESC and iPSC) No Stain PSCs typically in 90120 min Unconjugated primary antibodies, may be detected with secondary conjugated to fluorophore of choice Yes 100 g vial sufficient for staining four 24-well plates, twelve 6 cm dishes, or four 10 cm dishes
Product name Cat. No. Specificity Terminal assay Reaction time Color/label
ELF 97 Endogenous Phosphatase Detection Kit E6601 Medium (stains stem and progenitor cells) Yes Stains PSCs in 30 min Yellow-green fluorescent precipitate under Hoescht/DAPI longpass filter No 500 L vial sufficient for staining four 24-well plates, twelve 6 cm dishes, or four 10 cm dishes
FLoid protocols available Unit size
Yes 50 L vial sufficient for staining four 24-well plates, twelve 6 cm dishes, or four 10 cm dishes
Analyze
Assays
TaqMan Protein Assays TaqMan Protein Expression Assays are designed for accurate quantitation of protein expression using gold-standard TaqMan 5 nuclease chemistry with a workflow and sample quantity similar to our all TaqMan Gene Expression and microRNA (miRNA) Assays. As the protein expression results are obtained on the same analytical platform, these revolutionary assays enable direct correlation of mRNA and/or miRNA expression to protein expression. Learn more at lifetechnologies.com/taqmanprotein.
Selected instruments
Ion Proton Sequencer Genome and exome sequencing on your benchtop. Learn more at lifetechnologies.com/proton. QuantStudio 12K Flex Real-Time PCR System Delivering our highest throughput, flexibility, and scalability, the QuantStudio 12K Flex Real-Time PCR System is an all-in-one qPCR instrument that features five interchangeable blocks to support 96-well, Fast 96-well, 384-well, TaqMan Array Card, and OpenArray formats. Learn more at lifetechnologies.com/quantstudio.
67
Mesenchymal stem cells (MSCs)

Culture
We provide the broadest selection of Gibco cGMP compliant complete culture systems for mesenchymal stem cells (MSCs) and adipose-derived stem cells (ADSCs), some of which are free of animal-derived components. These media are designed to minimize adaptation time, maximize cell performance, and help you meet regulatory requirements.
MSC culture system selection guide.

The standard serum-based culture for MSCs and ADSCs Reduced-serum medium for lower variability and cost savings with serum-culture MesenPRO RS Medium 12746012 E071000 (New Zealand origin) IVD Multiple species; culture of cells sourced from bone marrow, adipose tissue, and umbilical cord Medium 1 x 500 mL MesenPRO RS Basal Medium 1 x 10 mL MesenPRO RS Growth Supplement GlutaMAX-I (Cat. No. 35050061) Fully documented serum-free MSC culture designed to reduce the burden in qualifying reagents and facilitate transition from bench to clinic StemPro MSC SFM CTS A1033201 Class II Medical Device Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and hair follicle Low 1 x 500 mL StemPro MSC SFM Basal Medium 1 x 75 mL StemPro SFM Supplement GlutaMAX-I CTS (Cat. No. A1286001) Substrate such as CELLstart CTS (Cat. No. A1014201) or Attachment Factor (Cat. No. S006100) Yes Stable at 4C for 4 weeks Yes
Product Cat. No. Product use Recommended applications
MSC-Qualified FBS 12662002, 12662011, 12662029 (USDA), 12664025 (Australia origin) 12763025 (US origin) IVD Multiple species; culture of cells sourced from bone marrow, adipose tissue, cord blood, umbilical cord, and placenta High 1 x 50 mL, 100 mL or 500 mL
Lot-to-lot variability Components/unit
Additional components required for complete medium
DMEM, low glucose (Cat. No. 11054020) GlutaMAX-I (Cat. No. 35050061) No Stable at 4C for 2 weeks No
Supports growth at high cell densityless medium, surface area, and time Stability of complete medium Specifically designed for cell therapy research
No Stable at 4C for 15 days No
MSC culture medium selection guide.

Serum-free, xeno-free MSC culture designed to reduce the risk and variability associated with animal-origin components Product Cat. No. Product use Recommended applications Lot-to-lot variability Components/unit Additional components required for complete medium Supports growth at high cell densityless medium, surface area, and time Stability of complete medium StemPro MSC SFM XenoFree A1067501 RUO Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and cord blood as well as pericytes and fibroblasts Low 1 x 500 mL StemPro MSC SFM Basal Medium 1 x 5 mL StemPro MSC SFM XenoFree supplement GlutaMAX-I CTS (Cat. No. A1286001) Substrate such as CELLstart CTS (Cat. No. A1014201) or Attachment Factor (Cat. No. S006100) Yes Stable at 4C for 2 weeks
68

MesenPRO RS Medium Cat. No. 12746012
A reduced-serum medium for MSC culture containing 2% FBS, MesenPRO RS Medium consistently improves expansion of MSCs compared with classical media (DMEM + 10% FBS) and maintains trilineage mesoderm differentiation potential.
StemProMSC SFM provides superior efficiency of human MSC expansion at high cell densities, requiring less medium, surface area, and time compared with classical medium (DMEM (low glucose) + 10% FBS).
StemPro MSC Serum-Free Medium (SFM) Cat. No. A1033201
StemPro MSC serum-free, xeno-free medium offers a completely animal originfree MSC culture system when used in conjunction with CELLstart CTS substrate. MSCs and adipose-derived stem cells (ADSCs) are grown in a more physiologically similar environment.
StemPro MSC SFM XenoFree Cat. No. A1067501
StemPro LipoMAX supplement is a human-derived, lipoprotein-based cell culture supplement for the improved expansion of MSCs and ADSCs.
StemPro LipoMAX Supplement Cat. No. A1085001
Xeno-free, fully defined cell culture substrate that contains components only of human origin. CELLstart CTS substrate enables attachment of MSCs and ADSCs.
CELLstart CTS Substrate Cat. No. A1014201
Cells
StemPro Human Adipose-Derived Stem Cell Kit Cat. No. R7788110
Human ADSCs isolated from human lipoaspirate tissue and cryopreserved from primary cultures. Kit contains human ADSCs and MesenPRO RS Medium.
These MSCs are isolated from the bone marrow of a C57BL/6 mouse at 8 weeks gestation.
Gibco Mouse (C57BL/6) Mesenchymal Stem Cells Cat. No. S1502100
These cells have a unique ability to track cells in transplantation and differentiation studies.
StemPro Rat Alk Phos Expressing Mesenchymal Stem Cells Cat. No. R7789120
Fluorescent visualization of StemPro Rat Alk Phos Expressing Mesenchymal Stem Cells using the ELF 97 Endogenous Phosphatase Kit.
69
Differentiation
When used in conjunction with StemPro MSC SFM or MesenPRO RS Medium, these kits provide a standardized culture workflow solution for MSC isolation, expansion, and differentiation into adipocytes, chondrocytes, and osteocytes. Learn more at lifetechnologies.com/mscdiff.
Gibco MSC differentiation kits
Complete differentiation into fat cells
StemPro Adipogenesis Differentiation Kit Cat. No. A1007001 StemPro Chondrogenesis Differentiation Kit Cat. No. A1007101
Complete differentiation into cartilage cells
Mesenchymal stem cells differentiated into fat cells and stained with Oil Red O, a fat-soluble dye.
Complete differentiation into bone cells
StemPro Osteogenesis Differentiation Kit Cat. No. A1007201
Analysis
The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSCs for laboratory investigations and preclinical studies, including specific surface antigen expression in which 95% of the cells express the antigens recognized by CD105, CD73, and CD90, with the same cells lacking (2% positive) the antigens recognized by CD45, CD34, CD14 or CD11b, CD79a or CD19, and HLA-DR (Cytotherapy 8:315 (2006)). Learn more about human MSC detection using the Attune Acoustic Focusing Cytometer at lifetechnologies.com/attune.
Human mesenchymal stem cells collected at passage 7 were identified as: (A) 99.7% CD73 +/0.29% CD19; (B) 99.4% CD73 +/0.22% CD45 ; (C) 99.7% CD90 +/0.09% CD34; (D) 99.9% CD90 +/0.07% CD14; (E) 99.2% CD105 +/0.76% HLA-DR. The data demonstrate that, using Gibco mesenchymal stem cell media, cell expression remains as expected after 7 passages.
70
Neural stem cells (NSCs)

Culture
Gibco media and supplements are the most widely cited reagents for use in neural stem cell culture. Choose between the flexibility of our B-27, N-2, StemPro Neural supplements and Neurobasal Medium or ease of use with the StemPro NSC SFM kit, specifically formulated for serum-free growth and expansion of human neural stem cells.
Designed for serum-free growth and expansion of human neural stem cells (hNSCs), with versatility to support both adherent and suspension NSC cultures.
StemPro NSC SFM Cat. No. A1050901
Serum-free media for the long-term viability of hippocampal and other neurons of the central nervous system. Neurobasal Medium is optimized for prenatal and embryonic neurons, while Neurobasal-A medium is best for growing postnatal and adult brain neurons. Learn more at lifetechnologies.com/neurobasal.
Neurobasal and Neurobasal-A Media
Natural Mouse Laminin Cat. No. 23017015
Supports the growth of human or rat neural stem cells, as well as neural primary cells.
Human Neural Stem Cells (H9-Derived) Cat. No. N7800100 Consistent, high-purity cells with normal human karyotypes that can differentiate into neurons, oligodendrocytes, and astrocytes. Consistent proliferation in adherent cell culture when used with StemPro NSC SFM.
These cells can be expanded as adherent or neurosphere suspension cell cultures while more than 75% of NSCs retain undifferentiated phenotype. Also supports differentiation into neurons, astrocytes, and oligodendrocytes.
Phenotype marker expressions of Rat Fetal Neural Stem Cells (proliferated up to passage 3) in StemPro NSC SFM medium. Cells retain their undifferentiated phenotype, and 90.9% cells express the NSC marker nestin.
Rat Fetal Neural Stem Cells Cat. No. N7744100
71
Differentiation
The following supplements are used in combination with a basal medium such as Neurobasal Medium.
B-27 Supplements are 50X liquid supplements for growth and long-term viability of hippocampal neurons. B-27 supplement is the most referenced supplement in neurobiology research, and is offered in both serum-free and xeno-free versions. Learn more at lifetechnologies.com/b27.
B-27 Supplements
N-2 Supplement is recommended for the growth and expression of neuroblastomas as well as post-mitotic neurons in primary cultures from both the peripheral nervous system (PNS) and the central nervous system (CNS).
N-2 Supplement Cat. No. 17502048
G-5 Supplement Cat. No. 17503012 G-5 Supplement is a 100X supplement recommended for growth and expression of glial cells (normal and tumor) of astrocytic phenotype (astrocytes).

Human embryonic stem cells differentiated into neurons.
72
Hematopoietic stem cells (HSCs)

Hematopoietic stem cells expressing CD34 (CD34+ cells) are defined by their ability to replenish all blood cell types and to self-renew. We offer CD34+ media, cells, and selection tools specifically formulated to support the culture of these stem cells.
Culture
Serum-free medium specifically formulated to support the development of human hematopoietic stem cells (CD34+) from sources including bone marrow, peripheral blood, and neonatal cord blood. It is manufactured with components of human, recombinant, and synthetic origin.
StemPro-34 SFM Cat. No. 10639011
Pooled Human Hematopoietic Progenitor cells (HPCs) derived from the umbilical cord blood pooled from multiple donors. Along with cord blood-derived CD34+ cells, this kit includes StemPro-34 SFM basal liquid medium and frozen StemPro-34 Nutrient Supplement to facilitate the immediate culturing of the StemPro CD34+ cells.
StemPro CD34+ Cell Kit Cat. No. A14059
Dynal CD34 Progenitor Cell Selection System Cat. No. 11301D
Stem cells can be isolated directly from whole blood, cord blood, or bone using this positive magnetic isolation of human CD34+ progenitor stem cells with bead release.
CD34+ cells, grown in StemPro-34 SFM, were reprogrammed into induced pluripotent stem cells using the CytoTune-iPS Sendai Reprogramming Kit.
73
Cell analysis
Cell analysis
Life Technologies offers a diverse array of reagents and instruments for cell analysis researchDynabeads cell isolation technology, Molecular Probes fluorescent products for monitoring cell function and health, cell structure, and cell tracing, and a broad portfolio of Molecular Probes and Novex primary and secondary antibodies for imaging and flow cytometry. We also offer a selection of new instruments for flow cytometry (Attune Acoustic Focusing Cytometer and Tali Image-Based Cytometer) and fluorescence imaging (FLoid Cell Imaging Station). The chart below highlights the most popular products and new technologies in this chapter. Follow the Web links provided in this chapter for a complete listing of all our products and services. If you have any questions about which of our solutions are right for you, please contact Life Technologies technical support by phone, email, Facebook at facebook.com/cytometry, facebook.com/immunologynews, or facebook.com/LIFEMolecularProbes, or Twitter at twitter.com/#!/MolProbes. Watch us on YouTube, too: youtube.com/lifetechnologies.
Section
Dynabeads cell isolation technology

Dynabeads cell isolation technology, page 76 Dynabeads CD3/CD28 technology, page 79
Cell function and health

Click-iT EdU, page 82 Premo FUCCI Cell Cycle Sensor, page 83 CellROX Deep Red Reagent, page 86
Probes for visualizing cell structure

CellLight fluorescent protein constructs, page 87
Key technologies
Cell isolation online selection guide, page 76 DynaMag magnets, page 80
LIVE/DEAD Viability/ Cytotoxicity Kits, page 81 CellEvent Caspase-3/7 Green Detection Reagent, page 85 pHrodo indicators, page 87
Alexa Fluor phalloidins, page 90 MitoTracker dyes, page 91 JC-1, page 91 LysoTracker dyes, page 92 BODIPY FL C5-ceramide, page 93 TO-PRO-3 dye, page 94
Everyday essentials Cell analysis
HulaMixer Sample Mixer, page 80
Human cell isolation, page 77 Mouse cell isolation, page 77 Positive, negative, and depletion isolation strategies, page 78 Mouse/human T cell activation and expansion, page 79
Cell viability and proliferation, page 81 Cell cycle, page 83 Apoptosis, page 84 Autophagy and reactive oxygen species, page 86 Endocytosis and phagocytosis, page 87 Intracellular ions, page 88
Cytoskeleton, page 90 Mitochondria, page 91 Lysosomes and peroxisomes, page 92 ER and Golgi apparatus, page 93 Nucleus, page 94
Applications
74
Cell analysis
HeLa cell labeled with CellLight Mitochondria-GFP (Cat. No. C10600) and CellLight Talin-RFP (Cat. No. C10612) reagents and with Hoechst 33342 nucleic acid stain (Cat. No. H21492).
Probes for cell tracing

Qtracker cell labeling, page 95, 96 FluoroMyelin Green Fluorescent Myelin Stain, page 98
Instruments
Attune Acoustic Focusing Cytometer, page 100
Primary antibodies for flow cytometry

Molecular Probes antibodies for flow cytometry, page 102 Qdot nanocrystals for multicolor flow cytometry, page 105
Secondary antibodies
Zenon antibody labeling, page 108 APEX antibody labeling, page 108
Calcein, AM, page 95, 96 CellTracker dyes, page 95 Fluorescent dextrans, page 95
Attune Acoustic Focusing Cytometer, page 100 FLoid Cell Imaging Station, page 101 Tali Image-Based Cytometer, page 99
Flow cytometer fluorophore selection guide, page 104 Primary antibody online selection guide, page 102
Alexa Fluor conjugated secondary antibodies and streptavidin, page 109 Secondary antibody online selection guide, page 109 Fluorescence SpectraViewer, page 111
Cell analysis
Cell tracking, page 95 Neuronal tracing, page 97
Fluorescence imaging, page 101 Flow cytometry, page 100, 102 Image-based cytometry, page 101
Simplified multiplexing options, page 103 Sample fixation and permeabilization, page 105
Immunodetection strategies, page 106 Create your own labeled antibodies, page 107 Labeled secondary antibodies, page 109 Choosing the right fluorophore, page 110 Imaging tools, page 110111
75
Cell analysis
Dynabeads cell isolation technology

Gentle, reproducible separation of your targeted cells
Viable and functionalproducts for positive isolation, nega Gentlecolumn-free separation and inert bead surfaces
translate to gentler handling of your cells and reduce the risk of contaminants in the preparations High yieldstube-based separation allows you to achieve excellent recovery of cells When cells are removed from their natural environment, there is a risk that experimental procedures will negatively impact cell phenotype and function. Choosing the right cell separation method is therefore critical to your downstream experiments. Dynabeads are superparamagnetic, monosized polymer beads coated with a thin, inert polymer shell to encase the magnetic material. This design helps to eliminate the risk that any unwanted material such as iron is left in the sample after separation. In addition, since the cells are never touched by the beads (negative-isolation and depletion beads) or are released from the beads after the gentle magnetic separation (positive-isolation beads), you get high purity and viability with no process-derived remnants that could affect your downstream results.
Dynabeads are uniform spherical beads with highly defined and consistent product characteristics, which helps ensure that you get truly reliable and reproducible results.
Dynabeads FlowComp Mouse CD4
600 500 400
tive isolation, cell activation, and depletion
Purity: 97.2%
Propidium iodide
105
104
Count
300 200 100 0
103
102
Viability: 86%
102 103 104 105
102
103
104
105
CD4+ Column-based isolation

400 350 300
CD4+
Propidium iodide
The table below organizes our cell isolation products according to cell origin and isolation method. If you go to lifetechnologies .com/cellisolation you can find information about our entire line of cell isolation products, and youll also have access to:
Purity: 78.5%
105
104
Count
250 200 150 100 50 0 102 103 104 105
103
Cell isolation and activation
Selection guides for choosing the correct cell Protocols for sample preparation and strategies for
cell isolation Data showing the performance of our cell isolation products versus other commercially available products Help in choosing the correct magnets for your tubes or plates Links to videos, brochures, and application notes and to references that cite the use of Dynabeads isolation product
102
Viability: 63%
102 103 104 105
CD4+
CD4+
Isolation of CD4+ T cells from mouse spleen cells. Cell isolation using Dynabeads FlowComp Mouse CD4 results in substantially higher purity (97%) and viability (86%) than column-based positive cell isolation (yielding purity and viability of 78% and 63%, respectively).
1,000 1,250
Count 750
500
250
102
103
104
105
500
Count 1,000
1,500
2,000
102
103
104
105
CD4 FITC
CD4 FITC
Purity of human CD4+ T cells. Purity before (left) and after (right) negative isolation from PBMC using Dynabeads Untouched Human CD4 T Cells.
76
Cell analysis
Human cell isolation
Gentle tube-based isolation of human cells directly from whole blood, MNC, buffy coat, bone marrow, or tissue samples for any downstream assay, including flow cytometry. Dynabeads are available for isolating human T cells, B cells, stem cells, NK cells, monocytes, dendritic cells, endothelial cells, tumor cells, leukocytes, granulocytes. Learn more at lifetechnologies.com/humancellisolation. Dynabeads are available for:
Incubate your starting sample with Dynabeads
Positive isolation and cell release Negative isolation resulting in untouched cells Depletion of unwanted cell types or positive cell isolation for Isolate cells using your own antibody
If you cant find a ready-to-use product for human cell isolation, we have a range of Dynabeads products that can be combined with an antibody of your choice to create a tailored cell isolation tool: Streptavidin Dynabeads, Secondary-coated Dynabeads, and Surface-activated Dynabeads. Learn more at lifetechnologies.com/humancellisolation. molecular applications
Separate bead-captured cells with a Dynal MPC magnet
Gentle tube-based isolation of mouse cells from directly from whole blood, spleen, lymph node, or thymus for any downstream assay, including flow cytometry. Dynabeads are available for isolating mouse T cells, B cells, NK cells, and dendritic cells. Learn more at lifetechnologies.com/mousecellisolation. Dynabeads are available for:
Negatively isolated cells (untouched) 1 2
Mouse cell isolation
Transfer supernatant with untouched cells to a new tube
Positive isolation and cell release Negative isolation of untouched cells Depletion of unwanted cell types or positive cell isolation for
molecular applications Isolate cells using your own antibody If you cant find a ready-to-use product for human cell isolation, we have a range of Dynabeads products that can be combined with an antibody of your choice to create a tailored cell isolation tool: Streptavidin Dynabeads, Secondary coated Dynabeads, and Surface activated Dynabeads. Learn more at lifetechnologies.com/mousecellisolation.
Positively isolated cells (bead-captured)
Rem
Cell-based assays, ow cytometry, cell expansion
ove b
eads
3
Molecular applications e.g., mRNA and DNA analysis
Workflow diagram for using Dynabeads for positive or negative tube-based cell isolation.
77
Cell analysis
Positive isolation
Positive-isolation kits use Dynabeads to isolate cells of interest directly from all types of sample material (e.g., human or mouse whole blood, bone marrow or peripheral blood mononuclear cells, spleen, or lymph nodes) by binding to antibody-coated Dynabeads. Positive isolation is ideal when cells of the highest purity and viability are required. Cells may be used for downstream analysis or as source material for the purification of RNA or proteins from the isolated fraction. FlowComp kits give you positive isolation of pure and viable cells directly from whole blood/buffy coat or PBMC. And, after a simple step in which the beads are removed, the cells can be used directly in all downstream experiments, including flow cytometry. Positive isolation:
Ideal for flow analysis and cell-based assays When maximum purity and yield is critical From whole blood, bone marrow, or buffy coat
Learn more at lifetechnologies.com/cellisolation.
Negative isolation
Negative isolation (Untouched) kits use Dynabeads in combination with an antibody cocktail to remove all unwanted cells from your sample, leaving only the target cells of interest behind. These kits are optimal for the isolation of fragile cells or when there are concerns that activation via antibody binding can interfere with downstream assays. Our antibody cocktails have been optimized to give you high cell purity and yield, and your resulting cells are both antibody- and bead-free. Negative isolation:
Truly untouched cells, bead- and antibody-free Cells in perfect shape for any application High yield, purity, and viability
Dynabeads depletion products are ideal for removal of one unwanted cell type at a time from your sample. Dynabeads are precoated with an antibody toward one target cell type that can be depleted from all types of starting materialeven viscous materials such as whole blood. Depletion:
Depletion
Easy removal of unwanted cell types Ideal for positive cell isolation for molecular applications High yield and purity
78
Cell analysis
Dynabeads for mouse and human T cell activation and expansion

Mimic in vivo activation of T cells
No need for antigen-presenting cells (APC) or antigen Gentle and easy, yet efficient, robust, and highly reproducible Activated cells retain their functional properties Use the same technology for mouse and human research-grade studies and
human clinical research The activation of naive and memory T cells is initiated through contact dependent antigen recognition involving antigen-presenting cells (APCs) such as dendritic cells. Activation occurs through the engagement of the T cell receptor complex (TCR/CD3, signal 1) and the CD28 costimulatory molecule (signal 2) on the surface of the T cell. Both signals are required to trigger cell-mediated immunity. Triggering of signal 1 alone can lead to T cell inactivation by anergy or apoptosis. Dynabeads T cell activation and expansion products offer a simple solution for mimicking the in vivo interaction of T cells with APCs by utilizing the two activation signals, and a threedimensional bead similar in size to APCs. Dynabeads CD3/CD28 can be used for both short-term activation and long-term expansion and are easily removed from culture. Activated cells are bead- and antibody-free, and their functions, cytokine profiles, and T cell repertoires reflect those of in vivo activation. Dynabeads CD3/CD28 technology is the only activation solution that allows for a controllable setting. Dynabeads CD3/CD28 technology is designed with covalently linked anti-CD3 and anti-CD28 antibodies for simple use and easy removal. Several CD3/CD28 antibody clones and ratios are available for more flexibility and optimization for your particular application. Custom conjugations are also available.
Nature
Dendritic cell
CD28
Resting T cell
Activated T cell
CD3/TCR
Expansion platform
Bead Anti-CD28
CD28
Resting T cell
Activated T cell
CD3/TCR
Anti-CD3
The Dynabeads T cell activation/expansion platform mimics in vivo activation/expansion.
Fold expansion
Dynabeads CD3/CD28/CD137 is specifically optimized for activation and expansion of mouse and human antigen-specific T cells. In addition to the anti-CD3 and anti-CD28 antibodies, the beads are also coated with anti-CD137 that recognizes CD137 (4-1BB), a costimulatory molecule expressed primarily but not exclusively on activated CD8+ and CD4+ T cells. Thus, Dynabeads precoated with monoclonal antibodies towards CD3, CD28, and CD137 specifically enhance proliferation of antigen-specific T cells. Learn more about Dynabeads for mouse and human T cell activation at lifetechnologies.com/cellactivation.
Dynabeads CD3/CD28/CD137 for antigen-specific T cell activation and expansion
3,500 3,000 2,500 2,000 1,500 1,000 500

2.4 4.7 10.9 20.5 446 50 190 865 1615 2970
10
11
12
Days
Expansion of naive T cells from human peripheral blood using Dynabeads Human T-Activator CD3/ CD28 (Cat. No. 11131D) for 12 days.
79
Cell analysis
Accessories for Dynabeads products

DynaMag magnets are optimized for efficient cell isolation using Dynabeads. The DynaMag magnets combine a strong magnetic attraction for efficient separations with flexible and smart ergonomic designs that ensure comfortable working positions. The magnets can be used for any starting sample in combination with all types of Dynabeads. We offer DynaMag magnets for these tube and plate configurations:
Get the highest utility and efficiency from Dynabeads products

DynaMag magnets
1.52 mL microcentrifuge tubes (straight tube racks and circular racks) 0.55 mL, 15 mL, and 50 mL tubes (Cat. Nos. 12303D, 12301D, 12302D) 6-well plates 24-well plates 96-well plates (side magnet and bottom magnet versions, skirted and half-skirted PCR strips (8 and 12 wells) 48- and 96-microtubes for the GeneAmp PCR System Closed, sterile blood bags (50330 mL static separations and >10 L in continuous
flow separations) Find specifications for all of these magnets online at lifetechnologies.com/magnets. plate versions, round- and flat-bottomed) (Cat. Nos. 12321D, 12320D)
DynaMag-5 Magnet (Cat. No. 12303D)
HulaMixer Sample Mixer
The HulaMixer Sample Mixer is perfect for sample preparation with Dynabeads products and for any other application that requires thorough mixing. The HulaMixer offers the following features:
Tilt, rotate, and/or vibrate your samples Continuous or timed operation All settings easily adjustable For use at 4C to 40C Two different platforms supplied to suit all kinds of tubes (0.550 mL)
See the HulaMixer Sample Mixer in action at lifetechnologies.com/hulamixer.
HulaMixer Sample Mixer (Cat. No. 15920D).
80
Cell analysis
Cell viability, proliferation, and health

Life Technologies offers a diverse selection of reagents for assessing cellular function and health. Many of the assays are fluorescence-based or colorimetric, offering sensitivity and convenience as well as safety. These products have been validated on multiple instrument platforms, including microscopy, flow cytometry, microplate readers, and high-content screening, enabling the analysis of a broad spectrum of cell functions:
Lymphocyte gate
SSC Viability gate
Viability and cytotoxicity Proliferation Apoptosis Autophagy Oxidative stress Endocytosis and phagocytosis Intracellular ions
Learn more about all of our cell function assays at lifetechnologies.com/cellfunction.
B
Live cells Dead cells 0.55%
LIVE/DEAD Viability/Cytotoxicity Kits
Simple to performadd, incubate, and analyze; no wash steps required Versatilecompatible with adherent cells, nonadherent cells, and certain tissues Simple to analyzegreen-fluorescent cells are live, red-fluorescent cells are dead
The LIVE/DEAD Viability/Cytotoxicity Kits provide an exceptionally easy fluorescencebased method for determining viability of adherent or nonadherent cells and for assaying cytotoxicity. LIVE/DEAD assays typically employ two separate fluorescence probes for clear live/dead distinction, and we have developed kits in several colors for flow cytometry (see right), microscopy (see below), or microplate formats. Learn more at lifetechnologies.com/livedeadkits.
CD4 Cy55PE
Fluorometric assays of cell viability and cytotoxicity are easy to perform with the use of a fluorescence microscope, fluorometer, fluorescence microplate reader, or flow cytometer, and they offer many advantages over traditional colorimetric and radioactivity-based assays. Our viability and cytotoxicity assay reagents and kits are principally used to enumerate the proportion of live and dead cells in a population.
Viability and cytotoxicity
Fixable Violet
CD8 Q705
Cell viability and health
CD4 Cy55PE
CD8 Q705
Live-cell gating with the LIVE/DEAD Fixable Violet Dead Cell Stain Kit (Cat. No. L34955) minimizes staining artifacts from analysis. In a comparison between live-cell gating using scatter (A), and livecell gating using LIVE/DEAD Fixable Violet dye (B), staining artifacts with scatter gating are illustrated. The significant number of dead cells in subsequent analysis using scatter (C) are noted, as compared to the use of the LIVE/DEAD Fixable Violet dye (D) to eliminate dead cells. Reproduced with permission from Elsevier (J Immunol Methods 313:199(2006)).
Fluorescence staining of live and dead cells with the LIVE/DEAD Cell Imaging Kit. HepG2 cells grown in 96-well microplate wells were stained with the LIVE/DEAD Cell Imaging Kit (Cat. No. R37601) and imaged on a Nikon Eclipse T200 microscope. Live cells are stained green; dead cells are stained red.
81
Cell analysis
SYTOX Dead Cell Stains
capabilities Ease of use: no wash steps; simply add, incubate, and analyze Fixable: Staining retained after fixation for simple live/dead analysis with intracellular phenotyping The cell-impermeant nucleic acid SYTOX Dead Cell Stains easily penetrate eukaryotic cells and both gram-positive and gram-negative bacteria with compromised plasma membranes, yet are completely excluded from live cells. Because they do not cross intact cell membranes and increase fluorescence upon double-stranded DNA binding, they are among our most brilliant dead cell stains. Easy to use, SYTOX dead cell stains can be applied to cells and visualized without an additional wash step because they are nonfluorescent in aqueous media. These stains can be used with multiple platforms including fluorescence microscopy, flow cytometry (see right), and microplates. See all colors of SYTOX Dead Cell Stains at lifetechnologies.com/sytox.
Number of cells
Accuracy: high-affinity nucleic acid stains for easy dead-cell discrimination Flexibility: multiple colors with minimal spectral overlap for expanded multicolor
1,000 800 600 400 200
0 101
102
103
104
105
106
SYTOX Green uorescence
Dead-cell discrimination using SYTOX Green Dead Cell Stain. A mixture of heat-killed and live Jurkat cells was stained with 30 nM SYTOX Green Dead Cell Stain (Cat. No. S34860) at room temperature for 20 min. Cells were analyzed on the Attune Acoustic Focusing Cytometer equipped with a 488 nm laser. Fluorescence emission was collected using a 530/30 nm bandpass filter. Live cells are easily distinguished from the brighter dead-cell population.
Cell proliferation and the characterization of agents that either promote or retard proliferation are extremely important areas of cell biology and drug-discovery research. We offer both traditional reagents for assessing cell proliferation and the cell cyclein particular the Hoechst nucleic acid stains and probes for 5-bromo-2-deoxyuridine (BrdU) incorporation during cell divisionas well as more advanced tools developed in our laboratories, including the Click-iT EdU cell proliferation assay. Click-iT EdU Cell Proliferation Assays
Cell proliferation and cell division
Accurate: superior to BrdU assays with minimal variation (low CVs) and much Speed: results in as little as 90 min Simplicity: streamlined protocol with fewer components than the original kits Versatility: assays for flow cytometry, imaging, HTS, HCS, and whole-animal
interrogations Click-iT EdU cell proliferation assay utilizes the power of click chemistry to provide a superior alternative to traditional methods for detecting and quantitating newly synthesized DNA. Click-iT assays use a modified nucleoside, EdU (5-ethynyl-2deoxyuridine), that is incorporated during DNA synthesis in a quick click chemistry reaction. The Click-iT EdU assay protocol is compatible with both adherent cells and cell suspensions. From start to finish, the EdU detection assay is complete in as little as 90 minutes, as compared with the antibody-based BrdU method, which takes 624 hours to complete. In addition, the Click-iT EdU cell proliferation assay can be multiplexed with surface and intracellular marker detection using Alexa Fluor dyelabeled secondary antibodies. Although the majority of applications are in cultured mammalian cells, Click-iT EdU reagents and methods have also been successfully applied to a wide range of model organisms. See all of the Click-iT assays at lifetechnologies.com/clickit. simpler protocol
Comparison of EdU and BrdU signal brightness. Sectioned formaldehyde-fixed, paraffin-embedded tissue was prepared from rats injected intraperitoneally with BrdU (upper panel) or EdU (lower panel). Proliferating cells within the intestinal villi that incorporated the nucleoside are pink. Nuclei in both samples were counterstained with Hoechst 33342 (gray). Images were acquired with a 20x objective on a Nikon Eclipse 800 epifluorescence microscope.
82
Cell analysis
CellTrace reagents for flow cytometry cell proliferation studies multiple generations
Number of cells counted
100
Superior performancebright single-peak staining enables visualization of Long-term signal stabilitywell retained in cells for several days post-staining Noncytotoxicno known effect on proliferative ability or biology of cells Versatilemultiple colors available to easily combine with antibodies or markers
of cell function, such as GFP CellTrace reagents allow you to permanently label cells to trace generations or divisions in vivo or in vitro without affecting morphology or physiology. We offer CellTrace reagents that are compatible with violet laser, 488 nm laser, and 633/635 nm laser excitation on your flow cytometer. See all of our CellTrace reagents at lifetechnologies.com/celltraceproliferation. Premo FUCCI Cell Cycle Sensor
101
102
103
104
CellTrace CFSE uorescence

Following cell proliferation in human peripheral blood lymphocytes using the CellTrace CFSE Cell Proliferation Kit (Cat. No. C34554). Human peripheral blood lymphocytes were harvested and stained with CellTrace CFSE (carboxyfluorescein diacetate, succinimidyl ester; 5(6)-CFDA, SE) on day 0. A portion of the population was arrested at the parent generation using mitomycin C (red peak). The remainder of the sample was stimulated with phytohemagglutinin and allowed to proliferate for 5 days. Solid green peaks represent successive generations.
Live-cell indicator of cell cycle status Flexible: works with live cells, but compatible with fixation Convenient: add to cells, incubate overnight, and visualize
The fluorescence ubiquitination cell cycle indicator (FUCCI) is a genetically encoded, two-color (red and green) indicator that allows you to follow cell division within a cell population (see below). We have incorporated the FUCCI genetic constructs into the powerful BacMam gene delivery systemthe Premo FUCCI Cell Cycle Sensor allowing a simple and efficient method for labeling cells and following their division. The Premo FUCCI Cell Cycle Sensor can be used to assess the effect of drugs, siRNA, or other factors on the transition of cells through the cell cycle. The fluorescence signals from geminin-GFP and Cdt1-RFP have been demonstrated to be resistant to fixation with 4% formaldehyde and permeabilization with 0.1% Triton X-100, thereby enabling processing of labeled cells with antibodies to other cellular targets. Learn more about the Premo FUCCI Cell Cycle Sensor at lifetechnologies.com/ fucci.
A
P)
M
Co lor le s
Green (ge mi nin -G F
-RF
P)
Yellow
Imaging cell cycle progression in live cells with the Premo FUCCI Cell Cycle Sensor.(A) Schematic of cell cycle progression with nuclear fluorescence changes. (B) U2OS cells were transduced with the Premo FUCCI Cell Cycle Sensor (Cat. No. P36237), then stained with Alexa Fluor 647 wheat germ agglutinin (Cat. No. W32466).
ed
(C
d t1
83
Cell analysis
Apoptosis
Apoptosis (programmed cell death) is the genetically controlled ablation of cells during normal development. Inappropriately regulated apoptosis is implicated in disease states such as Alzheimers disease, stroke, and cancer. In contrast to necrotic cells, apoptotic cells are characterized morphologically by compaction of the nuclear chromatin, shrinkage of the cytoplasm and production of membrane-bound apoptotic bodies. Biochemically, apoptosis is distinguished by fragmentation of the genome and cleavage or degradation of several cellular proteins. As with cell viability, no single parameter fully defines cell death in all systems; therefore, it is often advantageous to use several different approaches when studying apoptosis. Anticancer drug candidates failing to induce apoptosis are likely to have decreased clinical efficacy, making apoptosis assays important tools for high-throughput drug screening. Membrane asymmetry probe
Accurate apoptotic analysis on trypsinized cells Simple 5-minute staining protocol Compatible with other blue-excited apoptotic stains
The Violet Ratiometric Membrane Asymmetry Probe/Dead Cell Apoptosis Kit (Cat. No. A35137) provides an easy, efficient method for the detection of apoptosis with dead-cell discrimination using a violet laser flow cytometer (see figure, below). The Violet Ratiometric Membrane Asymmetry Probe detects the membrane asymmetry changes during apoptosis. It works well on adherent and suspension cells and correlates with other indicators of apoptosis, such as caspase detection and changes in mitochondrial membrane potential. The dye exhibits an excited-state intramolecular proton transfer (ESIPT) reaction resulting in dual fluorescence with two emission bands corresponding to 530 nm and 585 nm, producing a two-color ratiometric response to variations in surface charge. The F2N12S probe is combined with SYTOX AADvanced dead cell stain, which is capable of passing through the cell membrane only in late apoptotic or necrotic cells, allowing discrimination from early apoptotic cells. Unlike annexin-based assays, this assay does not require special buffers or wash steps, and it is less susceptible to the cell membrane damage commonly found during the physical or chemical removal steps when assaying adherent cells, therefore providing better data quality.
F2N12S green uorescence
196,608
A+D
F2N12S green uorescence
262,144
262,144
196,608
A+D
131,072
131,072
65,536
L
65,536
L
65,536
131,072
196,608
262,144
65,536
131,072
196,608
262,144
F2N12S orange uorescence
F2N12S orange uorescence
SYTOX AADvanced uorescence
105
D
SYTOX AADvanced uorescence
105
D
Violet ratiometric membrane asymmetry probe for apoptosis detection. Jurkat cells (T-cell leukemia, human) were treated with 10 M camptothecin for four hours (panels B and D) or left untreated as a control (panels A and C). Samples were analyzed on a flow cytometer with 405 nm excitation using 585 nm and 530 nm bandpass filters for F2N12S, and 488 nm excitation for SYTOX AADvanced dead cell stain using a 695 nm bandpass filter. Live cells can be discriminated from apoptotic and dead cells by the relative intensities of the two emission bands from F2N12S (A and B). In panels C and D, SYTOX AADvanced dead cell stain fluorescence is plotted against a derived ratio parameter from the two emission bands (585/530 nm) of F2N12S. A = apoptotic cells, L = live cells, D = dead cells.
104
104
103
103
A
102
102
101
1,250
2,500
3,750
5,000
101
1,250
2,500
3,750
5,000
F2N12S ratio 585 nm/530 nm uorescence x 1,000
F2N12S ratio 585 nm/530 nm uorescence x 1,000
84
Cell analysis
CellEvent Caspase-3/7 Green Detection Reagent
Optimized caspase-3/7 substrate for apoptosis analysis Simple, no-wash protocol helps preserve delicate apoptotic cells Compatible with both live-cell fluorescence imaging and formaldehyde-based fixation methods The CellEvent Caspase-3/7 Green Detection Reagent (Cat. No. C10423) is a novel fluorogenic substrate for activated caspase-3/7 that is compatible with both live-cell and fixed-cell imaging, with absorption/emission maxima of ~502/530 nm. Activation of caspase-3 is considered an essential event during apoptosis, making this an optimized reagent for analysis of apoptotic cells. The CellEvent Caspase-3/7 Green Detection Reagent is a fouramino acid peptide (DEVD) conjugated to a nucleic acid binding dye with an absorption/emission maxima of ~502/530 nm. The DEVD peptide sequence is a cleavage site for caspase-3/7, and the conjugated dye is nonfluorescent until cleaved from the peptide and bound to DNA. The CellEvent reagent is intrinsically nonfluorescent as the DEVD peptide inhibits the ability of the dye to bind to DNA. However, after activation of caspase-3/7 in apoptotic cells, the DEVD peptide is cleaved, enabling the dye to bind to DNA and produce a bright fluorogenic response. The fluorescence emission of the dye when bound to DNA is ~530 nm and can be observed using a standard FITC filter set. CellEvent reagent is ideal for imaging (see right), high-content screening, and flow cytometry. Click-iT TUNEL Alexa Fluor Imaging Assay
Multiplex imaging of apoptosis. U2OS cells were treated with 30 M etoposide for 18 hr to induce apoptosis. The treated cells were stained first with 7.5 M CellEvent Caspase-3/7 Green detection reagent (Cat. No. C10423, green fluorescence) to detect apoptosis and Hoechst 33342 nucleic acid stain (Cat. No. H3570, blue fluorescence) to label nuclei, and then with 150 nM MitoTracker Deep Red FM (Cat. No. M22426, pink fluorescence) to label mitochondria. Following fixation and permeabilization, actin was labeled with Alexa Fluor 546 phalloidin (Cat. No. A22283, orange fluorescence).
Robust workflow can be completed typically in 3 hours Available with Alexa Fluor 488, 594, and 647 dyes Much more sensitive than traditional antibody-based TUNEL methods Multiplexable with most other fluorescent reagents Ideal for imaging in vivo and in vitro, compatible with HCS and microplate assays
The Click-iT TUNEL assay (see right) is a breakthrough technology based on click chemistry for detection of DNA fragmentation. For TUNEL assays to yield meaningful results, it is necessary not only that the modified nucleotide is an acceptable substrate for TdT, but also that the detection method is sensitive and avoids detrimental loss of cells from the sample. See all of the Click-iT assays at lifetechnologies.com/clickit.
Late-stage apoptosis visualized using the Click-iT TUNEL Imaging Assay. HeLa cells were treated with staurosporine, then fixed and permeabilized. The Click-iT TUNEL Alexa Fluor 647 Imaging Assay (Cat. No. C10247) was performed. Activated caspase-3 was detected with a rabbit polyclonal primary antibody for cleaved caspase-3 and labeled with Alexa Fluor 488 goat antirabbit IgG antibody (green). Tubulin was detected with a mouse monoclonal anti-tubulin antibody and labeled with Alexa Fluor 555 goat antimouse IgG (orange). Nuclei were stained with Hoechst 33342 (blue). The light blue color represents an overlay of caspase (green), Hoechst (blue), and TUNEL (magenta) signals.
85
Cell analysis
Autophagy
A
Autophagy describes the segregation and delivery of cytoplasmic cargo, including proteins and organelles, for degradation by hydrolytic enzymes. The LC3B protein can be used as a general marker for autophagic membranes, and we developed the products below for imaging the LC3B protein and autophagic organelles in live cells. Premo Autophagy Sensors The Premo Autophagy Sensors (Cat. Nos. P36235 and P36236, see right) combine the selectivity of an LC3B-fluorescent protein chimera (GFP or RFP) with the transduction efficiency of BacMam technology, enabling unambiguous visualization of this protein in live cells. The reagent also tolerates fixation with formaldehyde and thus is compatible with fixed-cell analysis, which is required for multiplex experiments using antibodies, and is preferred for HCS analysis workflows. To image autophagy, simply add BacMam LC3-FP to cells and incubate overnight for protein expression. LC3B Antibody Kit The LC3 Antibody Kit for Autophagy (Cat. No. L10382) includes a highly specific rabbit polyclonal antibody against LC3B that has been validated for use in fluorescence microscopy and high-content imaging. Also included is a control compound for inducing autophagosomesfollowing treatment with this compound, lysosomal pH increases and the normal autophagic flux is disrupted, resulting in autophagosome accumulation. Learn more about LC3B protein detection at lifetechnologies.com/autophagy.
Expression of the Premo Autophagy Sensor LC3BGFP. (A) Imaging of Premo Autophagy Sensor in rat hippocampal neurons. (B) Imaging of Premo Autophagy Sensor (green) and CellLight LysosomesRFP (red) in HeLa cells.
Oxidative stress
CellROX Deep Red Reagent
Compatible with fluorescent imaging, florescent plate readers, flow cytometry, and Highly resistant to photobleaching Multiplexible: compatible with GFP and Alexa Fluor 488 dye Versatile: suitable for live-cell imaging or fixed samples
high-content screening
CellROX oxidative stress reagentsincluding CellROX Green reagent (Cat. No. C10444), CellROX Orange reagent (Cat. No. C10443), CellROX Deep Red reagent (Cat. No. C10422), and a variety pack (Cat. No. C10448)are fluorogenic probes designed to reliably measure reactive oxygen species (ROS) in live cells. The cellpermeable CellROX reagents become brightly fluorescent upon oxidation, with excitation/emission maxima at 485/520 nm, 545/565 nm and 640/665 nm, respectively. The CellROX Green reagent binds to DNA (fluorescent signal is localized primarily in the nucleus and mitochondria). In contrast, CellROX Orange and Deep Red reagents localize to the cytoplasm. The resulting fluorescence can be measured using traditional fluorescence microscopy, high-content imaging and analysis, microplate fluorometry, or flow cytometry. The staining workflow is simple, and the reagent can be applied to cells in complete growth medium or buffer. All of the CellROX reagents are very photostable when compared with traditional ROS detection dyes. In addition, both CellROX Green and CellROX Deep Red reagents are retained in cells after formaldehyde fixation, providing assay flexibility and improved workflows when compared with classic dyes used for ROS detection. CellROX Green staining is also stable to detergent permeabilization.
Imaging oxidative stress with CellROX Deep Red reagent. (A) Human osteosarcoma (U2OS) cells were treated with 100 M menadione for 1 hr to induce oxidative stress, and then stained with 5 M CellROX Deep Red reagent, 5 g/mL of CellMask Orange plasma membrane stain, and 1 M SYTO Green fluorescent nuclear stain for 30 min at 37C. The cells were washed 3 times with HBSS before imaging.
86
Cell analysis
Endocytosis and phagocytosis
The pHrodo dye and CellLight reagents are fluorescence-based tools that can be used to interrogate routes of molecules and microbeswhether critical to cell health or pathogenicacross the plasma membrane, as well as their final destination after entering the cell. pHrodo indicators The proprietary pH-sensitive pHrodo dye is a specific sensor of endocytosis. The dye is nonfluorescent at neutral pH and fluoresces bright red in acidic environments (see right). This increase in fluorescence signal at low pH makes it ideal for studying endocytosis and its regulation by drugs and/or environmental factors. For multiplexing, pHrodo dye is available in red and green colors, and the signal can be preserved by formaldehyde fixative. pHrodo dye is available as convenient conjugates of E. coli, S. aureus, and Zymosan A BioParticles for phagocytosis and dextran BioParticles for endocytosis. If you want to make your own conjugations, both pHrodo green and red are available in various reactive chemistries and in antibody labeling kits. The pHrodo dye allows you to:
Discriminate endocytosis from adherent and extracellular particles Specifically detect phagocytosis and endocytosis with a fluorogenic dye Reduce signal variability and improve timing in sensitive experiments Can be multiplexed with other fluorescent dyes
Learn more about all of our pHrodo indicators at lifetechnologies.com/phrodo.
CellLight reagents CellLight reagents label a number of intracellular organelles (see below), including endosomes, with either GFP or RFP. CellLight reagents use fluorescent proteins that survive fixation and permeabilization, and therefore they can be used in combination with any antibody. Labeling of transferrin, epidermal growth factor, or low-density lipoprotein with Alexa Fluor dyes provides a useful means of visualizing ligand internalization. Some of our more popular fluorescent probes are also available in 0.1 mL format, providing a cost-effective way for researchers to try these innovative products.
Time-lapse images showing internalization and acidification of the pHrodo E.coli BioParticles conjugate during phagocytosis by Murine J774A.1 cells. Using excitation with a 561 nm laser, fluorescence images were recorded in the 570699 nm emission range and white-light DIC images were recorded on a separate PMT. Images were collected every 30 sec for 83.8 min (only the first and last images in the time course are shown here) using a Leica TCS SP5 laser confocal microscope employing a 63 (NA 1.4) oil objective and the resonant galvanometer scanner mode (8000 Hz). Image contributed by Lucy Deriy and Deborah Nelson, University of Chicago. See Nelson Lab time-lapse video of this process.
Human osteosarcoma (U2OS) cells expressing CellLight Tubulin-GFP (Cat. No. C10509) were treated with 200 M tert-Butyl hydroperoxide for 2 hr. A stain solution containing 5 M CellROX Orange (Cat. No. C10443), and 2 drops/ml of NucBlue Live Cell Stain (Cat. No. R37605) was applied for 30 min at 37C. Cells were washed and imaged with Live Cell Imaging Solution (Cat. No. A14291DJ).
Human osteosarcoma (U2OS) cells expressing CellLight Actin-RFP were treated with 100 M menadione for 1 hr. A stain solution containing 5 M CellROX Green (Cat. No. C10444) and 2 drops/mL of NucBlue Live Cell Stain (Cat. No. R37605) was applied for 30 min at 37C. Cells were washed and imaged with Live Cell Imaging Solution (Cat. No. A14291DJ).
87
Cell analysis
Intracellular ions
A
Ion channels provide exciting opportunities for drug development, targeting a diverse array of therapeutic areas including neurological, cardiovascular, and pulmonary diseases, pain, and inflammation. To address the most challenging ion channel targets in any desired cell background, researchers need an assortment of optimized assays for their discovery efforts. Our product offering for cell-based ion channel assays is designed for sensitivity, ease-of-use, and assay flexibility. Learn more at lifetechnologies.com/ionchannelassays. Cell-based ion channel assays.
Ion Potassium Assay technology FluxOR Potassium Ion Channel Assay Description Functional, HTS-validated universal potassium assay (Cat. Nos. F10016, F10017) lifetechnologies.com/fluxor BacMam Ion Channel Targets BacMam-mediated delivery of potassium ion channels including hERG lifetechnologies.com/bacmamionchannels Calcium Fluo-4 Direct Calcium Assay Kit Add-and-read, homogenous fluorescent calcium readout validated for HTS (Cat. Nos. F10471, F10472, F01473) lifetechnologies.com/fluo4direct Premo Cameleon Calcium Sensor Fluorescent proteinbased calcium biosensor protein lifetechnologies.com/premocameleon Chloride Premo Halide Sensor Voltage sensor probes Fluorescent proteinbased chloride biosensor protein lifetechnologies.com/premohalide Membrane potential FRET-based, ratiometric assessment of membrane potential lifetechnologies.com/voltagesensorprobes
Dose-dependent calcium response to muscarnic 1 (M1) receptor agonists and antagonists. CHO M1 cells were plated in a poly-D-lysine coated 384-well plate and incubated overnight. The following day, cells were assayed for a calcium response to carbachol using the Fluo-4 Direct Calcium Assay Kit. Cells were stimulated with agonists, carbachol, MCN-A-343, bethanechol, oxotremorine, and pilocarpine (A) or cells were treated with antagonists, scopolamine, telenzipine, and DAMP to block the calcium response elicited by 114 nM carbachol (EC80 for this receptor) (B). Measurements are given in relative fluorescent units (RFU) as the maximum response minus the minimum response divided by the minimum response. Rank order of agonist and antagonist potency agreed with published results.
The TC-FlAsH II and TC-ReAsH II detection kits (Cat. Nos. T34561, T34562) and Gateway Expression Vectors (Cat. No. T34563) offer a tag-based fluorescence labeling technology for live cell imaging and more. The FlAsH-EDT2 and ReAsH-EDT2 labeling reagents become fluorescent when they bind to recombinant proteins containing the tetracysteine (TC) motif Cys-Cys-Pro-GlyCys-Cys. FlAsH and ReAsH technology provides sensitive live-cell imaging and subcellular localization of proteins containing the TC-tag using fluorescence microscopy. The TC-FlAsH II and TC-ReAsH II detection kits are ideal for protein localization or real-time protein production studies, though its versatility offers a range of benefits: A B
FLAsH and ReAsH tetracysteine-based protein detection
Rapid protein detectioneasily constructed six-amino acid tag Multiplexing flexibilitychoice of red or green fluorescence from the same Small tag sizereduces interference with target protein Stable but non-covalent bindingallows dual labeling, pulse-chase experiments
and other dynamic, real-time studies Compatible with live- or fixed-cell imaging and multiplexing Multiple applications beyond direct labeling for imaging Affinity purification Learn more at lifetechnologies.com/flashreash. tagged protein
CHO-K1 cells expressing a tetracysteine-tagged version of -tubulin, labeled with TC-ReAsH reagent. Upon treatment with vinblastine, a compound known to perturb cytoskeletal structure, tubulin drastically rearranges from a reticular structure (A) to rod shapes (B).
88
Cell analysis
Probes for visualizing cell structure

This section highlights our most popular and easiest-to-use fluorescent dyes and fluorescent protein-based reagents for staining organelles and subcellular structures. Additional information is available online; go to lifetechnologies.com/ organellestainsselectionguide to find selection guides for over 100 different fluorescent probes, organized by these cellular structures:
Information about related products, including organelle-specific antibodies A link to our interactive Cell Staining Simulation Tool that allows you to select Access to immunofluorescence protocols, the Fluorescence Spectra Viewer, the
Image Gallery, and more compatible sets of fluorescent dyes for your experiment
Adiposomes Cytoplasm Cytoskeleton Endoplasmic reticulum Golgi complex Intracellular membranes Lysosomes Membrane trafficking Mitochondria Nuclear envelope Nucleoli Nucleus Peroxisomes Plasma membrane Whole-cell stains for image segmentation in HCS applications
Cell structure
Nucleus
Nucleolus Golgi complex
Tubulin
Lysosome
Endoplasmic reticulum
Mitochondria Cytosol
Peroxisome
Actin
Lipid rafts
Plasma membrane
89
Cell analysis
The cytoskeleton is an essential component of a cells structure and one of the easiest to label with fluorescent reagents. We offer labeling reagents for monomeric actin (G-actin), filamentous actin (F-actin), tubulin, and other cytoskeletal proteins. Read more about the options for detecting actin, intermediate filaments, and tubulin in our BioProbes magazine article at lifetechnologies.com/actintubulindetection. Find a more detailed version of the selection guide below at lifetechnologies.com/ organellestainsselectionguide.
Product Alexa Fluor dyelabeled phalloidins Popular products (fluorescence color) A22281 (blue), A12379 (green), A34055 (orange), A12381 (orange), A34054 (far red), A22287 (far red), A22286 (near IR) Key attributes Alexa Fluor dyelabeled phalloidins bind to F-actin in nanomolar concentrations and show equal affinity for large and small filaments; phalloidins are typically used to stain fixed cells; live-cell staining requires liposome delivery or microinjection of conjugates, compromised membranes, or unlabeled phalloidins; contrast between stained and unstained areas is extremely large; phalloidin-labeled actin remains functional. Learn more about phalloidin conjugates for staining actin at lifetechnologies.com/phalloidinconjugates. Rhodamine phalloidin R415 (orange) Conjugated to a popular orange-fluorescent dye, tetramethylrhodamine (TRITC); convenient probe for labeling, identifying, and quantitating F-actin in formaldehyde-fixed and permeabilized tissue sections, cell cultures, or cell-free experiments; rhodamine phalloidin will stain live cells. Fusions of fluorescent proteins to the N-terminus of -actin in mammalian cells; allow visualization of cytoskeletal components in live cells or cytoskeletal rearrangements in HCS cytotoxicity assays. Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts. CellLight Tubulin-GFP, CellLight Tubulin-RFP C10613 (green), C10614 (red) Fusions of fluorescent proteins to the N-terminus of -tubulin that allow live-cell imaging of microtubules; to monitor tubulin changes as an indicator of cytotoxicity; also for following cell division in transiently transduced cells, including primary and stem cells. Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts.
Cytoskeleton
CellLight Actin GFP
C10582 (green)
Cell structure
CellLight MAP4-GFP, CellLight MAP4-RFP
C10598 (green), C10599 (red)
Fusions of fluorescent proteins to microtubule associated protein 4 (MAP4), enabling visualization of microtubules in live cells that can also readily be fixed for endpoint studies such as HCS-based cytotoxicity analyses. Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts.
TubulinTracker Green
T34075 (green)
Results in green-fluorescent staining of polymerized tubulin in live cells.
Live bovine pulmonary arterial endothelial cells were labeled with CellMask Deep Red plasma membrane stain (Cat. No. C10046), the tubulin-selective TubulinTracker Green dye (Oregon Green 488 Taxol, bis-acetate, Cat. No. T34075), mitochondrion-selective MitoTracker Red CMXRos dye (Cat. No. M7512), and bluefluorescent Hoechst 33342 nuclear stain (Cat. No. H21492).
Muntjac skin fibroblasts stained with Alexa Fluor 488 phalloidin, (Cat. No. A12379) an antia-tubulin antibody (Cat. No. A11126) pre-labeled with Zenon Alexa Fluor 568 Mouse IgG1 Labeling Kit (Cat. No. Z25006) and the anticdc6 antibody pre-labeled with Zenon Alexa Fluor 350 Mouse IgG1 Labeling Kit (Cat. No. Z25000).
The actin cytoskeleton of the Muntjac fibroblast was visualized using the green-fluorescent Alexa Fluor 488 phalloidin (Cat. No. A12379). The nucleus was stained with TO-PRO-3 reagent (Cat. No. T3605).
90
Cell analysis
Mitochondria are found in eukaryotic cells, where they make up as much as 10% of the cell volume. They are pleomorphic organelles with structural variations depending on cell type, cell-cycle stage, and intracellular metabolic state. The key function of mitochondria is energy production through oxidative phosphorylation and lipid oxidation. We offer these probes for live-cell staining of mitochondria. Find out more at about these products and about anti-OxPhos complex antibodies for fixed cell staining at lifetechnologies.com/organellestainsselectionguide.
Product MitoTracker Green FM Popular products (fluorescence color) M7514 (green) Key attributes Live-cell stain; staining does not appear driven by mitochondrial function. Useful counterstain for mitochondrial mass. Has been reported to exhibit specific staining after fixation in certain cells. Good live-cell stains, and well suited for multicolor labeling experiments; all are well retained after fixation except MitoTracker Red FM.
Mitochondria
MitoTracker Red FM, MitoTracker Orange CMTMRos, MitoTracker Red CMXRos, MitoTracker Deep Red FM MitoTracker Orange CM-H2TMRos, MitoTracker Red CM-H2XRos CellLight MitochondriaGFP, CellLight MitochondriaRFP
M7510 (orange), M22425 (red), M7512 (red), M22426 (deep red)
M7511 (orange), M7513 (red)
Do not fluoresce until they enter live cells, where they are oxidized to the fluorescent mitochondrion-selective probe MitoTracker Red CM-H and then sequestered in mitochondria. Fluorescent proteins targeted specifically to mitochondria via the leader sequence of E1 pyruvate dehydrogenase; delivered to live cells using BacMam delivery technology. Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts. Widely used cationic dye that is dependent upon mitochondrial membrane potential for accumulation; JC-1 monomer (low concentration) is green, whereas accumulation at higher concentration shifts emission to the red, allowing ratiometric membrane potential measurements. Contains MitoTracker Red CMXRos dye and Hoechst 33342 for highly selective mitochondrial and nuclear staining in live GFP-transfected cells. This indicator is a cationic derivative of dihydroethidum and has been designed for highly selective detection of superoxide in the mitochondria of live cells.
C10508 (green), C10505 (orange)
JC-1
T3168 (green as monomer, red as aggregate)
Image-iT LIVE Mitochondrial and Nuclear Labeling Kit MitoSOX Red Indicator
I34154 (contains two stains: blue and red fluorescent) M36008 (red)
Cell structure
Bovine pulmonary artery endothelial cell (BPAEC). MitoTracker Red CMXRos (Cat. No. M7512), SYTOX Green nucleic acid stain (Cat. No. S7020), biotin-XX goat antimouse IgG antibody and Cascade Blue NeutrAvidin biotin-binding protein (Cat. No. A2663).
Bovine pulmonary artery endothelial cell (BPAEC). Oregon Green 514 goat antimouse IgG antibody, (Cat. No. O6383) MitoTracker Red CMXRos (Cat. No. M7512) and DAPI (Cat. No. D1306).
HeLa cell labeled with CellLight Mitochondria-GFP (Cat. No. C10600) and CellLight Talin-RFP (Cat. No. C10612) reagents and with Hoechst 33342 nucleic acid stain (Cat. No. H21492).
91
Cell analysis
Lysosomes and peroxisomes
Lysosomes are single membrane-bound vesicles that contain digestive enzymes such as glycosidases, acid phosphatases, elastase, cathepsins, carboxypeptidases and a variety of other proteases. The function of this organelle is to break down waste products and cellular debris. Peroxisomes are membrane-bound organelles in the cytoplasm of the cell. They are responsible for removing harmful peroxides from the cell and also play a role in fatty acid metabolism. We offer these probes for live-cell staining of lysosomes and for live- and fixed-cell staining of peroxisomes. Find out more about these products and about antibodies to lysosome and peroxisome proteins for fixed-cell staining at lifetechnologies.com/organellestainsselectionguide.
Product LysoTracker Blue DND-22 Popular products (fluorescence color) L7525 (blue) Key attributes Highly selective for acidic organelles at nanomolar concentrations, shows minimal staining of mitochondria, for labeling and tracking of acidic organelles in live cells, loads rapidly. Is taken up very rapidly by live cells; imaging using this dye is best between 1 and 5 minutes after dye is added. Rapid loading, highly selective for acidic organelles at nanomolar concentrations, show minimal staining of mitochondria, for labeling and tracking of acidic organelles in live cells. LysoSensor dyes are live-cell stains that are nonfluorescent at pH 7 and exhibit a pH-dependent increase in fluorescence upon acidification. Contains LysoTracker Red DND-99 dye and Hoechst 33342 for highly selective staining of lysosomes and nucleus in live GFP-transfected cells. Fluorescent proteins targeted to lysosomal associated membrane protein 1; for live-cell visualization of lysosomes and any process in which they are involvedphagocytosis, endocytosis, autophagy. Learn more about CellLight reagents at lifetechnologies.com/ celllightlabelingproducts. CellLight Peroxisome-GFP C10604 (green) Fluorescent protein targeted specifically to the peroxisome via the peroxisomal C-terminal targeting sequence; for live-cell staining. Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts.
LysoTracker Green DND-26 LysoTracker Red DND-99
L7526 (green) L7528 (red)
LysoSensor Blue DND-167, LysoSensor Green DND-153 Image-iT LIVE Lysosomal and Nuclear Labeling Kit
L7533 (blue), L7534 (green)
I34202 (contains two stains: blue and red fluorescent)
CellLight Lysosomes-GFP, C10596 (green), C10597 (red) CellLight Lysosomes-RFP
Cell structure
SelectFX Alexa Fluor 488 Peroxisome Labeling Kit
S34201 (green)
Kit for peroxisome labeling in fixed cells, providing a primary antibody directed against peroxisomal membrane protein 70 (PMP 70), an Alexa Fluor 488labeled secondary antibody, PBS, blocking solution, fixative, and permeabilization solutions.
Live bovine pulmonary artery endothelial cells (BPAEC) stained with LysoTracker Red DND-99 (Cat. No. L7528). Then, a solution of green fluorescent dihydrorhodamine 123 (Cat. No. D632, D23806) and blue-fluorescent Hoechst 33258 (Cat.No. H21491) was added and allowed to incubate with the cells for an additional 10 minutes before the cells were subsequently washed and visualized.
Imaging autophagy in live HeLa cells with CellLight reagents for mitochondria and lysosomes. Cells were transduced with CellLight Lysosomes-GFP (Cat. No. C10596) and CellLight Mitochondria-RFP (Cat. No. C10601). Following treatment with chloroquine to induce autophagosome accumulation, nuclei were stained with Hoechst 33342 (Cat. No. H21492) and live-cell imaging was performed.
92
Cell analysis
The endoplasmic reticulum (ER) and Golgi apparatus are primarily responsible for the proper sorting of lipids and proteins in cells. Consequently, most of the cell-permeant probes for these organelles are either lipids or chemicals that affect protein movement. The flattened membranous sacs of the ER and the Golgi apparatus can be stained with a variety of lipophilic probes and then distinguished by their morphology. Find out more about these products and about antibodies to endoplasmic reticulum and Golgi apparatus proteins for fixed-cell staining at lifetechnologies.com/organellestainsselectionguide.
Product ER-Tracker Green, ER-Tracker Red CellLight ER-GFP, CellLight ER-RFP Popular products (fluorescence color) E34251 (green), E34250 (red) Key attributes Live-cell permeant and highly endoplasmic reticulum-selective BODIPY dyes; when cells are stained using the provided protocol, staining patterns are partially retained after aldehyde fixation. Fluorescent proteins targeted specifically to the nucleus via the endoplasmic reticulum organelle signal sequence of calreticulin and KDEL (ER retention signal). Learn more about CellLight reagents at lifetechnologies.com/ celllightlabelingproducts. SelectFX Alexa Fluor 488 ER Labeling Kit S34200 (green) Kit for endoplasmic reticulum organelle labeling in fixed cells, providing a primary antibody directed against ER-associated disulfide isomerase (PDI), an Alexa Fluor 488 dye-labeled secondary antibody, PBS, blocking solution, fixative and permeabilization solutions. Fixed cell stain for ER staining; concanavalin A selectively binds to -mannopyranosyl and -glucopyranosyl residues. Selective staining of the Golgi complex with applications for lipid metabolism and trafficking studies; brighter, more photostable than NBD conjugates; concentration-dependent shift from green to red in the Golgi complex; good for outlining cell boundaries for observation of morphogenetic movements and for measuring rates of lipid synthesis by Schwann cells. Selective staining of the Golgi complex with applications for lipid trafficking studies, brighter and more photostable than the NBD conjugates; displays no concentration-dependent emission. Lectins selectively bind to terminal -N-acetylgalactosaminyl residuesan intermediate sugar added in O-linkage to serine and threonine residues in cis-Golgi cisternae and then substituted with galactose and sialic acid in the trans-Golgi organelle; for each cell type or tissue it is necessary to empirically determine appropriate lectins and staining conditions. Fluorescent proteins targeted specifically to the Golgi complex via the human Golgi-resident enzyme N-acetylgalactosaminyltransferase-2. Learn more about CellLight reagents at lifetechnologies.com/celllightlabelingproducts. NBD C6-ceramide N1154 (green) Classic structural marker for the Golgi complex.
Endoplasmic reticulum and Golgi apparatus
Concanavalin A Alexa Fluor 594 conjugate BODIPY FL C5-ceramide, complexed to BSA
C11253 (red) D3521, B22650 (green)
BODIPY TR ceramide, complexed to BSA Lectin HPA Alexa Fluor 488 conjugate
D7540, B34400 (red)
L11271 (green), L32454 (red)
Cell structure
CellLight Golgi-GFP, CellLight Golgi-RFP
Organelle staining of live bovine pulmonary artery endothelial cells. Endoplasmic reticulum was labeled with ER-Tracker Green (Cat. No. E34251); mitochondria were visualized with MitoTracker Red CMXRos (Cat. No. M7512).
U2OS osteosarcoma cells labeled with CellLight ER-GFP reagent (Cat. No. C10590) and with Hoechst 33342 nucleic acid stain (Cat. No. H21492).
93
Cell analysis
Nucleus
The nucleus of the cell is responsible for regulation of gene expression and is a membrane bound organelle that is comprised of the nuclear envelope, nucleoli, and nuclear lamina. Find out more about these products and about antibodies to nuclear proteins for fixed cell staining at lifetechnologies.com/organellestainsselectionguide.
Product NucBlue Live Cell Stain (Hoechst 33342 Special Formulation) NucBlue Fixed Cell Stain (DAPI Special Formulation) 7-AAD Ethidium homodimer-1 Popular products (fluorescence color) R37605 (blue) Key attributes Ready-to-use liquid DAPI formulation in a dropper bottle; excellent cell permeable nuclear counterstain for live cells. Ready-to-use liquid DAPI formulation in a dropper bottle; excellent nuclear counterstain for fixed cells. Nuclear stain, GC-selective, weakly permeant in some cell types. One of the highest-affinity fluorescent probes available for nucleic acid staining; binds to dsDNA, ssDNA, RNA, and oligonucleotides with a significant fluorescence enhancement (>40-fold); the dyes intercalation is not sequence-selective. One of the most commonly used fixed-cell nuclear stains. Fluorescent proteins targeted specifically to nucleus via the SV40 nuclear localization sequence; allows visualization of the nucleus without utilizing DNA-binding dyes; can also be used as a segmentation marker in endpoint HCS studies. Learn more about CellLight reagents at lifetechnologies.com/ celllightlabelingproducts. Alternatives to DNA staining for localizing the nucleus and chromosomes; allow localization of histones and following of cell division in live cells. Learn more about CellLight reagents at lifetechnologies.com/ celllightlabelingproducts. SelectFX Nuclear Labeling S33025 (several) Kit, for fixed cells (DAPI, SYTOX Green, 7-AAD, TO-PRO-3 iodide) A sampler kit of our most popular nuclear stains for fixed cells, with optimized protocols for use.
R37606 (blue) A1310 (red) E1169 (red)
Propidium iodide Organelle Lights Nuc-GFP, CellLight Nucleus-CFP, CellLight Nucleus-GFP, CellLight Nucleus-RFP
P3566 (red) O36209 (green), C10616 (blue), C10602 (green), C10603 (red)
CellLight Histone 2B-GFP, C10594 (green), C10595 (red) CellLight Histone 2B-RFP
Cell structure
SYTO 9, SYTO 14, SYTO 82, SYTO 59, SYTO 61
S34854 (green), S7576 (green), S11363 (orange), S11341 (red), S11343 (red)
Live-cell nuclear counterstains; permeant to eukaryotic and prokaryotic cell membranes; SYTO 9 is useful to indicate bacterial viability; SYTO 61 stains live and dead bacteria; SYTO 14 is twice as fluorescent when complexed with RNA as with DNA. Highest-affinity SYTOX stain, fixed-cell stain, useful as a dead-cell stain, 500x increase in fluorescence upon nucleic acid binding, can be used in conjunction with blue and red stains for multiplexing. Fixed-cell nuclear stain, some mitochondrial staining, shorter-wavelength emission than propidium iodide, useful as a dead-cell stain, 500x increase in fluorescence upon nucleic acid binding, can be used in conjunction with blue and green stains for multiplexing. Ultrasensitive dsDNA detection, dead-cell stain, electrophoresis prestain, used as an alternative to annexin conjugates for analyzing apoptotic cells.
SYTOX Green nucleic acid stain
S7020 (green)
SYTOX Orange nucleic acid S11368 (orange) stain
TO-PRO-3 iodide
T3605 (far red)
U2OS osteosarcoma cells transduced with CellLight Histone 2B-RFP (Cat. No. C10595) and CellLight MAP4-GFP (Cat. No. C10598) reagents.
94
Cell analysis
Probes for cell tracking and neuronal tracing

Molecular Probes fluorescent cell trackers and neuronal tracers are essential tools for following cells in complex environments, and have been used extensively in studies of cell migration, wound healing, and stem cell differentiation. These probes (Table 1) are stable, nontoxic at working concentrations, well retained in cells (from hours to days), and brightly fluorescent at physiological pH. Moreover, they are available in fluorescence emission colors that span the spectrum, allowing simultaneous detection of cell movement and other critical cell functions. Learn more about all of our fluorescent cell trackers and neuronal tracers at lifetechnologies.com/celltracing. Table 1. Molecular Probes fluorescent cell tracking probes and tracers.
Probe class
Calcein AM probes
Retention time
<1 day (13hr)
Fixable General attributes

No Acetoxymethyl (AM) estermodified compounds that can passively diffuse across cell membranes, where nonspecific esterases cleave the AM ester moieties to produce a product that is retained only in live cells. Baculovirus-packaged DNA constructs for the expression of nontargeted fluorescent proteins or of fluorescent proteins fused to signal peptides for specific targeting to key cytoskeletal structures and organelles. Cell-permeant, thiol-reactive labeling reagents containing a chloromethyl moiety that can react with intracellular thiol groups from cysteine residues in proteins or glutathione. Cell-permeant, amine-reactive labeling reagents containing a succinimidyl ester that forms stable adducts with primary amines located intra- and extracellularly. Provides Qdot probes that contain a selective targeting peptide for uptake into the cytoplasm of live cultured cells. Once inside cells, Qdot probes provide intense, stable fluorescence that can be traced through several generations. Highly water-soluble dyes and other detectable probes that can be used as cell tracers. In most cases, the polarity of these probes is too high to permit them to passively diffuse through cell membranes. Consequently, special methods for loading the dyes into cells must be employed, including microinjection, pinocytosis, or techniques that temporarily permeabilize the cell membrane.
Instrument platform * Selected Cat. Nos.

FC, FM For imaging: C3099, C3100MP, C1430, C34852, For flow cytometry: C34858, C34853, C34851 B10383, C10582, C10601
CellLight reagents
Days to weeks or Yes 68 generations
FC, FM, HCS
CellTracker 68 days or probes 68 generations CellTrace probes Qtracker Cell Labeling Kits Polar tracers
Yes
FC, FM, HCS FC, FM
C7025, C2925, C34552, C2927 C34557, C34554, C34553
510 days or Yes 810 generations Days to weeks or Yes 810 generations
FC, FM, HCS, M
Q25021MP, Q25041MP, Q25071MP, Q25001MP,
Hours to days
Some
FM
A12924, A12923, A12922, A10436, A20501MP, A10436, A20502, B1592
Cell tracing
Membrane tracers
Hours to weeks
Some
FM, FC Lipophilic dialkylcarbocyanines, including DiI, DiO, DiD, and DiR, are widely used retrograde and anterograde tracers in live and fixed tissues. Ready-to-use NeuroTrace DiI and DiO tissue-labeling pastes can be applied directly to tissues using the tip of a needle, allowing dye penetration into bundled neurons for labeling axons both on and below the surface. Also available are the NeuroTrace Multicolor Tissue-Labeling Kit, which contains DiI, DiO, and DiD pastes, and the Lipophilic Tracer Sampler Kit, which contains DiI, DiO, DiD, DiR, and DiA analogs. Dextrans are hydrophilic polysaccharides characterized by their moderate to high molecular weight, good water solubility, and low toxicity. Polystyrene fluorescent microspheres that can be used for a wide range of applications including blood flow determination, tracing, in vivo imaging, and calibration of imaging and flow cytometry instruments. Cholera toxin B conjugate is a powerful tool for retrograde labeling of neurons. Other protein conjugates, such as albumin and ovalbumin, are used to target receptors, detect protease activity, and measure intracellular pH. Nissl staining is a standard histochemical method for visualizing neurons in the brain and spinal cord. FM, FC
D282, D3899, D3911, D7756, N22880, D275, D3898, N22881, D307, D7757, D12731
Fluorescent dextran conjugates
Hours to weeks
Yes
D22910, D1956, D3308, D1830, D22914, D7139, D22913 F8760, F8770, F8787, F21012, F20892, F13083 C22841, C34775, C22842, C22843, C34776, C34778, A13100, A23015 N21479, N21480, N21481, N21482, N21483
Fluorescent Hours to days microspheres Proteins & protein conjugates NeuroTrace Nissl stains Hours to weeks
Some
FM, FC
Yes
FM, FC, HCS
Days to weeks
No
FM
* FC = flow cytometry. FM = fluorescence microscopy. HCS = high-content screening and analysis. M = microplate assay. After fixation, these emGFP and tagRFP fusions can be detected withanti-GFP antibodiesandanti-RFP antibodies, respectively. Lysine-fixable dextrans contain lysines and can therefore be fixed in place with formaldehyde or glutaraldehyde. Fixable dextrans contain free amines (but not lysines) and can be fixed in place with formaldehyde or glutaraldehyde.
95
Cell analysis
Calcein AM probes
The traditional nonfixable calcein probes are useful tracers for both cell tracing and viability studies. These tracers contain acetoxymethyl (AM) moieties that block the inherent negative charges on the molecules, allowing their passage across cell membranes. Once inside the cell, nonspecific esterases cleave the AM esters to produce cell-impermeant polar molecules that can be efficiently retained in the cell for several hours. Unlike the CellTracker and CellTrace probes described below, the Calcein AM probes are retained only in live cells. When paired with a cell-impermeant nucleic acid stain such as SYTOX Green dye or propidium iodide, these calcein tracers are effective for differentiating viable and dead cells using microscopy (see figure, right) or flow cytometry. Calcein probes are easy to load and are effective for 2-8 hours, depending on the cell type.
Calcein AM. Jurkat cells were incubated with either green-fluorescent calcein AM (C1430, C3099, C3100MP), CellTrace calcein red-orange AM (C34851), or calcein blue AM (C1429). The labeled cells were then combined and imaged with the appropriate filters.
24 hr post-wound with 0.1 m cytochalasin D
Control
24 hr post-wound
Cell tracing
CellTracker fluorescent probes (see figure, right) freely pass through cell membranes and are converted to cell-impermeant reaction products. The cell-impermeant form is passed to daughter cells through several generations but is not transferred to adjacent cells in the population. Cells that are loaded with CellTracker probes are typically fluorescent and viable for at least 24 hours, making these probes excellent long-term cell tracers. Furthermore, the staining pattern can be fixed with formaldehyde or glutaraldehyde to allow long-term storage of the labeled cells or tissue, to facilitate safe handling of potentially pathogenic samples, and to permit signal amplification in cases where the anti-dye antibody is available. CellTracker probes are not transferred to adjacent cells in a population, and are ideal long-term tracers for transplanted cells or tissues. The signal from CellTracker probes is typically monitored using fluorescence microscopy, and the probes are offered in several fluorescent colors for compatibility with multicolor analyses.
CellTracker probes
Wound-healing assay with CellTracker Green CMFDA. (Left) After growing in culture for 72 hr, a monolayer of Gibco human neonatal dermal fibroblasts was stained with CellTracker Green CMFDA. (Middle) The stained cell monolayer was scratched and left to heal for 24 hr. Cells have reinvaded the wound but are not as dense as in the control plate. (Right) The stained cell monolayer was scratched and then treated with 0.1 M cytochalasin D for 24 hr to disrupt actin polymerization, resulting in no closure of the wound site.
The CellTrace probes are also well retained in cells and are retained after formaldehyde-based fixation once they have reacted with proteins or other amine-containing biomolecules. These amine-reactive cell tracersincluding CellTrace Violet, CellTrace CFSE, and CellTrace Far-Red DDAO-SE probeseach contain a succinimidyl ester that reacts with available primary amines located both inside and outside the cell. The bright, homogeneous staining of CellTrace Violet probe shows very little fluorescence variation between cells in a population, allowing visualization of distinct generations of proliferating cells in a fluorescence histogram.
CellTrace probes
Qtracker probes. (A) Qtracker 625. (B) Three-color Qtracker cell labeling of 3T3 (green), HeLa (red), and U188 (white) cells labeled with Qtracker 565, 655, and 705, respectively. (C) Vascular labeling with Qtracker vascular labels.
extended time periods. These Qdot nanocrystals contain a targeting peptide that allows their selective uptake into the cell, where they are distributed in vesicles throughout the cytoplasm (see figure, above). Inside the cell, the Qdot nanocrystals exhibit an intense, photostable fluorescence that can be observed using continuous illumination, without time constraints due to photobleaching or degradation. Their fluorescence is maintained in complex cellular environments and under various biological conditions, including changes in intracellular pH, temperature, and metabolic activity, and they are passed to daughter cells through at least 7 generations, or about 8 days in U2OS cells. Qtracker Cell Labeling Kits have been used for diverse applications including in vivo tracking of hematopoietic stem cells, following
The Qtracker Cell Labeling Kitsavailable with Qdot nanocrystals in seven brilliant colorscontain the reagents needed to deliver fluorescent Qdot nanocrystals into live cells, providing a powerful method for real-time tracking studies over
Qtracker probes
96
Cell analysis
Schwann cells after injection into nerve grafts, and detecting neutrophil survival in infected skin wounds. For in vivo vascular imaging in small animals, Qtracker nontargeted quantum dots are designed to be injected into the tail vein of mice for the study of vascular structure using small animal in vivo imaging (SAIVI) techniques (see figure). These nanocrystals exhibit intense fluorescence with red-shifted emission for increased tissue penetration and avoidance of background autofluorescence, and have a PEG surface coating developed to minimize nonspecific interactions and reduce immune response. Because the PEG surface coating does not contain reactive functional groups, the Qtracker non-targeted quantum dots are retained in circulation longer and can be imaged for up to 3 hours with a single injection, or for longer periods of time with additional injections. tracers, which evenly distribute throughout the whole cell, are good tools for total neuronal cell visualization (see figure, below). These dyes can also be used to visualize gap junctions. Membrane tracers Lipophilic dialkylcarbocyanines, including DiI (see figure, below), DiO, DiD, DiR, FM 1-43, and FM 4-64, can be introduced into membranes by direct application or with kits (Vybrant celllabeling solutions). Lateral diffusion of these dyes eventually stains the entire cell. These probes are widely used as retrograde and anterograde tracers in live and fixed tissues, as well as for long-term assays of cellcell association, and are available in a wide selection of fluorescence emission colors and physical forms (solid, dissolved in organic solvents, suspended in paste, and large crystals). Ready-to-use NeuroTrace DiI and DiO tissue-labeling pastes can be applied directly to tissues using the tip of a needle, allowing dye penetration into bundled neurons for labeling axons both on and below the surface. Also available are the NeuroTrace Multicolor Tissue-Labeling Kit, which contains DiI, DiO, and DiD pastes, and the Lipophilic Tracer Sampler Kit, which contains DiI, DiO, DiD, DiR, and DiA analogs.
CellLight reagents are ready-to-use fusion constructs of emGFP and TagRFP for the expression of untargeted, cytoskeleton-targeted, or organelle-targeted fluorescent proteins. The CellLight cytoplasmic markeralso called BacMam GFP transduction controlproduces emGFP expression in the cytosol and nucleus of a wide range of cell types. The transiently transduced cells typically express the fluorescent protein fusion for about 5 days in standard cell lines (e.g., HeLa and CHO). With slowly dividing cellssuch as some primary cellsexpression has been demonstrated for up to 2 weeks, and with terminally differentiated neurons we have observed expression for more than 3 weeks after transduction. Learn more about CellLight reagents at lifetechnologies.com/ celllightlabelingproducts.
CellLight reagents
Neuronal tracing
Fixable polar tracers We prepare a wide variety of highly water-soluble dyes and other detectable probes that can be used as cell tracers. Molecular Probes fixable polar tracers include the membraneimpermeant probes lucifer yellow and Alexa Fluor hydrazides. In most cases, the polarity of these water-soluble probes is too high to permit them to passively diffuse through cell membranes. Consequently, special methods for loading the dyes into cells must be employed, including microinjection, pinocytosis or techniques that temporarily permeabilize the cell membrane. These probes are often microinjected, and are useful for a variety of applications including neuronal tracing, investigating cellcell and cellliposome fusion, membrane permeability, transport through gap junctions, and cell uptake during pinocytosis. These fluorescent hydrazide derivatives are covalently linked to surrounding biomolecules during aldehyde fixation, allowing subsequent antibody-based analyses. Lucifer yellow and sulforhodamine 101 (Orange/Red) are widely used tracers that must be loaded by injection. These polar
Cultured left-upper quadrant neurons from the sea slug Aplysia californica that have been microinjected with either lucifer yellow CH (L453, L682, L1177, L12926) or sulforhodamine 101 (S359). Image contributed by David Kleinfeld, AT&T Bell Laboratories, and Brian Salzberg, University of Pennsylvania School of Medicine.
Cell tracing
DiI, the classic red membrane stain. DiI is nonfluorescent in aqueous solutions, but has incredibly intense fluorescence in membranes. In this image, DiI (D282, D3911) is used as a diagnostic tool to evaluate patterns of innervation in a newborn mouse cochlea. The larger image is of a mutant cochlea and the inset is of a wildtype cochlea. Images contributed by Bernd Fritzsch, Creighton University, and L. Reichardt and I. Farinas, Howard Hughes Medical Institute, San Francisco.
97
Cell analysis
Fluorescent dextran conjugates Fluorescent dextran conjugates (such as Alexa Fluor dextrans) are ideal cell-lineage tracers because they are relatively inert, exhibit low toxicity, and are retained in cells for long periods. Dextrans are offered in a range of molecular weights and are useful size-exclusion probes for determining pore sizes in membranes. They are typically loaded into cells by microinjection, whole-cell patch clamping, or electroporation.
Stains and kits for neuronal tracing Nissl staining is a standard histological method for retrograde tracing of neurons in the brain and spinal cord. Composed of ribosomal RNA associated with the rough endoplasmic reticulum in neuronal perikarya and dendrites, the Nissl substance redistributes within the cell body in injured or regenerating neurons, providing a marker for the physiological state of the neuron. Our NeuroTrace Nissl stains are selective for the Nissl substance characteristic of neurons, and provides more sensitivity than traditional histological dyes like toluidine blue or cresyl violet. These Nissl stains are available in a wide spectrum of fluorescent colors for staining neurons, either alone or in combination with immunofluorescence staining of specific proteins. The BrainStain Imaging Kit (see figure, below) enables 3-color combinatorial labeling of myelin, neurons, and nuclei in brain cryosections in a single 20-minute staining step plus washes. This kit contains novel stains that can be used separately or together in one staining solution, replacing traditional methods that can take 13 days. Standard histochemical methods such as immunohistochemistry are compatible with these stains. Each BrainStain kit provides:
Fluorescent and biotinylated dextrans are routinely employed to trace neuronal projections. Dextrans can function efficiently as anterograde or retrograde tracers, depending on the study method and tissue type used. Active transport of dextrans occurs only in live, not fixed tissue. Dextran conjugates with molecular weights up to 70,000 daltons have been employed as neuronal tracers in a wide variety of species. Fluorescent microspheres FluoSpheres and TransFluoSpheres polystyrene microspheres are intensely fluorescent, durable, and inert tracers. These microspheres are particularly useful as long-term markers for transplantation studies. They are either injected into cells or taken up by phagocytosis. These microspheres satisfy several prerequisites of ideal long-term biological tracers. Because the dyes in our microspheres are incorporated throughout the microsphere rather than just on its surface, the fluorescence output per microsphere is significantly greater than that obtained from protein or dextran conjugates and is relatively immune to photobleaching and other environmentdependent effects. FluoSpheres and TransFluoSpheres microspheres are also biologically inert and physically durable, and they are available with a large number of uniform sizes and surface properties. Furthermore, their spectral properties can be freely manipulated during manufacture without altering their surface properties. Proteins and protein conjugates Protein tracers have molecular weights between ~12,000 (cholera toxin subunit B conjugates) and ~240,000 (B- and R-phycoerythrin). Molecular Probes cholera toxin conjugates are made from a recombinant version of the B subunit only. This allows us to provide a high-purity product completely free of the toxic A subunit. Cholera toxin B subunit (CT-B) attaches to cells by binding to ganglioside GM1, making it a powerful tool for retrograde labeling of neurons. This tracer has been used in a variety of applications, including tracing of rat forebrain afferents, projections of the parabrachial region, and neurons of the urinary bladder wall. When used in neuronal tracing applications, CT-B is typically introduced by pressure injection or by iontophoretic injection into neural tissue. Other protein conjugates, such as albumin and ovalbumin, are used to target receptors, detect protease activity, and measure intracellular pH.
FluoroMyelin Green fluorescent myelin stain NeuroTrace 530/615 red-fluorescent Nissl stain DAPI nuclear counterstain
For more details about Molecular Probes fluorescent cell tracers, visit lifetechnologies.com/celltracing.
Cell tracing
BrainStain Imaging Kit. Mouse brain cryosections of the hippocampal region (top) and diencephalon region (bottom), labeled simultaneously with three dyes: DAPI (nuclei, blue), FluoroMyelin Green stain (myelin, green), and NeuroTrace 530/615 red Nissl stain (neuronal cell bodies, red).
98
Cell analysis
Tali Image-Based Cytometer

Visualize, quantify, and analyze your cells
Versatilegenerate visual and analytical data on GFP/RFP expression, apoptosis, and cell viability Convenientset up and analyze cells in minutes right at your bench, with minimal cleaning and maintenance Accurateobtain statistically significant 3-parameter population analysis The Tali Image-Based Cytometer is a 3-channel benchtop imaging platform that performs quantitative analysis of GFP/RFP expression, apoptosis, cell viability, and much moreright at your bench. And, its fast, delivering a typical cell population analysis in about 45 seconds using only 25 L of sample. The Tali cytometer contains bright-field, red fluorescence, and green fluorescence channels, enabling researchers to simultaneously count green- and/or red-fluorescent stained cells, as well as cells expressing GFP and RFP. The Tali cytometer displays fluorescence data for the population in histograms, which can be adjusted (gated) to collect the fluorescence of interest. Both the raw cell-by-cell data and simple reports can then be exported for archiving and sample comparisons. The Tali Image-Based Cytometer is ideal for:
The Tali Image-Based Cytometer
Rapid analysis of GFP/RFP vector transfection efficiency Identification of GFP- and/or RFP-positive cells (see figure below) Simple two-color apoptosis assays using annexin V and PI (see figure below) Accurate population analysis of cell viability Quick verification of samples before sorting or flow cytometry analysis
Find the Tali Image-Based Cytometer, consumables, reagents, accessories, and application data at lifetechnologies.com/tali.
Instruments and accessories
Simultaneous analysis of GFP- and RFP-expressing cells. The Tali Image-Based Cytometer displays green and red fluorescence channels, enabling simultaneous quantitation of green and red fluorescence from proteins and stains. The Tali cytometer also provides actual cell counts and automatically calculates cell concentration for each cell population.
The Tali Apoptosis Assay. The Tali Apoptosis Assay uses an annexin VAlexa Fluor 488 conjugate and propidium iodide to assess cell viability and health. Results indicate whether each cell is live (unstained), dead (yellow and red), or apoptotic (green).
99
Cell analysis
Attune Acoustic Focusing Cytometer

The real answer at the speed of sound
Better data in less time with less effort:
Faster acquisitionmore events/second and faster run time/sample Better sensitivity and precisionlow CVs even at high collection rates Simplified sample preparationno-lyse, no-wash method eliminates cell loss Easy to usefree and intuitive software allows you to analyze data from Low cost of ownershiplow fluid consumption and direct count without
counting beads The Attune Acoustic Focusing Cytometer, the worlds first and only acoustic focusing flow cytometer, uses ultrasonic waves to align cells into a single focused line. With the Attune Acoustic Focusing Cytometer, you get both high sample input rate and high precision. You control the sample concentration, the flow rate, the number of photons you detect, the length of your experiment, and more, which allows you to maximize the data from each cell sample. But best of all, the Attune cytometer makes it easy with streamlined sample prep and easy-to-use acquisition and analysis software. Powered by Molecular Probes flow reagents, Attune cytometer offers two laser configurations (blue/red or blue/violet), and the Attune Autosampler and enables you to optimize performance and throughput without sacrificing data quality.
SSC
Attune Acoustic Focusing Cytometer
and improves data accuracy anywhere at any time
8 7 6 5 4 3 2 1
Attune sample rate: 1,000 L/min
Research applications supported
The Attune Acoustic Focusing Cytometer has been used to investigate aspects of cancer biology, stem cells, oceanography, and microbiology. In fact, cells up to 100 m in diameter have been run without clogging our 200 m flow cell, and the instrument also handles particles as small as 1 m. Most standard applications for flow cytometry have been tested on the Attune cytometer, including: Apoptosis Rare-event detection Basic phenotyping (up to 6 colors) Phagocytosis Cell cycle analysis Detection of phosphoproteins Cell proliferation assays Detection of intracellular markers Live/dead cell discrimination
CD34 102 103
CD34+
0 101
104
105
106
No software licensing fees
Identification of CD34+ cells from peripheral blood. Peripheral blood from a normal donor was stained and run on the Attune Acoustic Focusing Cytometer at a collection rate of 1,000 L/min with a stop gate set at 500,000 total cells. A rare population (0.045%) of CD34+ cells (red box) was identified within the population of cells with an acquisition time of 4 min 28 sec.
The Attune Cytometric Software offers limited copies and can be downloaded without licensing fees. You can analyze the results from any computer at your convenience at any time from anywhere, and an intuitive user interface eases the learning curve.
Simple, user-defined analysis Automated or user-defined compensation Automated experimental settings that can be completely customized and saved for Automated instrument checks Recover unused samples (tube only)
Learn about the advantages of acoustic focusing technology and the Attune cytometer at lifetechnologies.com/attune. future experiments
100
Cell analysis New Molecular Probes reageNts FLoid Cell Imaging Station siMPlify fluoresceNt iMagiNg
Click. Print. Love. Yes, its that easy
Perfect for a quick check of your stained cells, the FLoid Cell Imaging Station is designed to deliver high-quality, 3-color fluorescent cell images right at your benchtop with an interface so simple even novice users can collect data in just a few clicks of the mouse.
Simplified protocols
Simplecollect images in no time with the intuitive user interface Practicalprint cell images and place them in your notebook Accessiblecapture fluorescent cell images at your bench, not in the darkroom Robustprotect expensive confocal microscopes from excessive day-to-day use Informativefind the most popular Molecular Probes reagents
Meet FLoid at lifetechnologies.com/floid.
Ready to use with no dilutions or calculations
Convenient packaging for easy storage

B
The FLoid Cell Imaging Station
backDropbackgroundsuppressor
Effectively suppresses background fluorescence for live cell imaging No pipetting, calculations, or dilutions Place right on your bench where you need it Immediate reduction of blue, green or red background signal
ReadyProbes reagents
Focus on your data, not reagent preparation with new ReadyProbes imaging reagents
Quickuser-friendly dropper-bottle formulations with minimal Intuitiveclear, concise, streamlined protocols Accessibleroom temperaturestable reagents
BPAE Molecular cells labeled with CellLight Golgi-RFP in DMEM, 20% FBS without Find Probes ReadyProbes reagents (A) and with (B) BackDrop Background Suppressor at lifetechnologies.com/readyprobes.
or no dilutions
image-itfixation/PermeabilizationKit
No need to prepare fresh fixative Room temperature storage
Complete, ready to use kit to fix, permeabilize and wash your
Compatible with fluorescent proteins, dyes, antibodies, and iT reagents
labeling BPAE with and red-fluorescent Texas Red-X phalloidin for BPAE cells cellsstained were fixed permeabilized using the Image-iT F-actin, mouse monoclonal anti-tubulin in conjunction with green-fluorescent FixationPermeabilization Kit. BODIPY FL goat antimouse IgG for labeling microtubules, and blue-fluorescent DAPI for labeling the nuclei. Image was captured using the FLoid Cell Imaging Station. Prior to immunostaining, cells were fixed and permeabilized with the readyto-use Image-iT Fixation/Permeabilization Kit (Cat. No. R37602).
101
Cell analysis
Your solution for multicolor flow cytometric analysis

We offer a diverse array of highly specific RUO, ASR, and IVD primary antibodies to targets studied in both basic and clinical research settings. These antibodies are validated for flow cytometry and include specificities for CD markers, cytokines, chemokines, and growth factors, as well as antibodies with utility for oncology, immunology, cell signaling, apoptosis, proliferation, and stem cell research. From subset identification of heterologous cell populations to rare-event detection, multicolor flow cytometry can help answer complex cell biological questionsin less time, with less sample.
Flexiblebroadest range of dye, protein, and Qdot nanocrystal antibody conjugates Compatiblewide breadth of specificities for numerous types of applications Reliablequality products for clinical and research applications, RUO, ASR, and IVD Trustedreferred to in over 20,000 journal citations
reagents available
We offer 24 different fluorescent dyes (including the Alexa Fluor family of dyes, Pacific Blue and Pacific Orange dyes), proteins (PE, PerCP, and APC), and Qdot nanocrystals conjugated to a wide range of antibodies, including ten different tandem conjugates to further expand your multiparametric flow cytometry capabilities. Go to lifetechnologies.com/flowantibodies to find:
Product details for over 950 antibodies validated for use in flow cytometry, including Our online Flow Cytometry Resource Center (lifetechnologies.com/flowresources) that
allows you to access our video tutorials, webinars, protocols, links to published literature and more Our web-based Primary Antibody Selection Tool that allows you to search for antibodies by protein target, species reactivity, host, label, and application A downloadable Fluorophores for Flow Cytometry Selection Guide to help you find dyes that are compatible with 20 different commercially available cytometers A link to our Flow Cytometry mobile app human, mouse, rat, and monkey specificities
102
Cell analysis
Simplify multiplexing with our broad color selection
The availability of more color options allows more efficient experiments, gaining more information with less sample, in less time. Through multiplexing, experimental error is diminished by minimizing the number of samples used to obtain results. Our wide selection of fluorophores allows researchers to select panels with minimal spectral overlap.
250 200
Qdot 655-CD3 (655/20)
Qdot 655-CD3 (655/20)
Qdot 605-CD4 (605/20)
105
1.7%
105
0.0%
105
0.0%
Side scatter
104
4.5%
104
0.9%
104
0.0%
150 100 50
102
103
103
0 -533
0 -533
0 -529
22 10 10
1033 10
4 104
1055 10
Qdot 705-CD45 (710/30)
-629
103
104
105
Qdot 605-CD4 (605/20)
-807
103
104
105
APC-Alexa Fluor 750-CD8 (780/60)
-807
103
104
105
Fluorescein-CD2 (525/50)
Qdot 655-CD3 (655/20)
Qdot 605-CD4 (605/20)
Qdot 605-CD3 (605/20)
Qdot 655-CD3 (655/20)
105
0.0%
105
6.9%
105
6.7%
105
1.0%
104
0.0%
104
1.3%
104
1.5%
104
59.6%
103
103
103
103
0 -629 -807
0 -629
0
-633
Gated: lymphocyte + monocyte
-633 -4,882 -103 0 103 104 105
103
104
105
APC-Alexa Fluor 750-CD8 (780/60)
-1,969
103
104
105
RPE-CD16+CD56 (575/26)
RPE-Cy7-CD19 (780/60)
-265
0 102
103
104
105
APC-CD14 (670/14)
Human PBLs were labeled with anti-CD4 Qdot 605, anti-CD3 Qdot 655, anti-CD45 Qdot 705, anti-CD2 FITC, anti-CD16+CD56 RPE, anti-CD19 RPE-Cy7, anti-CD14 APC, and anti-CD8 APC-Alexa Fluor 750. Samples were run on an LSR II flow cytometer. Plots are gated on lymphocytes by side scatter and CD45, except as noted. Axes are labeled with the filters used; plots are labeled with compensation values.
103
Cell analysis
Fluorophores to fit every common laser and instrument configuration
By leveraging years of expertise in the creation and optimization of various technologies including organic dyes, fluorescent proteins, and Qdot nanocrystals, we are able to offer a complete portfolio of Molecular Probes fluorescent labels that spans the near-UV, visible, and near-IR spectrum. Regardless of your instrument and its various on-board excitation sources, we have labels designed to help you get the most out of every sample and every flow cytometry run.
Fluorophore selection guide for flow cytometry.

Laser Fluorophore Alexa Fluor 405
UV
Violet
488 nm
532 nm
561 nm
633/635 nm
Emission (nm) 421 455 551
Pacific Blue Pacific Orange Qdot 605
Qdot 655 Qdot 705
Qdot 800 Alexa Fluor 488
Fluorescein PerCP R-phycoerythrin (R-PE, PE) PE-Texas Red PE-Alexa Fluor 610 TRI-COLOR (TC, PE-Cy5)
605 655
705 800 519 525
678 575 615 628 670 694 723 767 660 668 694 696 775 695
PE-Cy5.5 PE-Alexa Fluor 700 PE-Cy7 Allophycocyanin (APC) Alexa Fluor 647 APC-Cy5.5 Alexa Fluor 700
APC-Alexa Fluor 750 PerCP-Cy 5.5
PE-Alexa Fluor 700
723
104
Cell analysis
Qdot nanocrystals for multicolor flow cytometry
Qdot nanocrystals are nanometer-scale semiconductor particles with unique fluorescence properties that make them attractive labels for multicolor flow cytometry. These fluorophores are optimally excited by a UV or violet (405407 nm) laser, but can also be excited with longer-wavelength light sources. Qdot nanocrystals are brighter than most organic fluorophores when compared to conventional dyes. And unlike conventional dyes that photobleach over time, Qdot conjugates remain fluorescent under constant illumination. This fluorescence stability allows ample time for flow cytometric analysis and permits additional analysis steps after sorting. With Qdot nanocrystals you get:
Multiplexing capabilityadd Qdot nanocrystal conjugates to existing marker experiments to increase StabilityQdot nanocrystals do not degrade over time like tandem conjugates, which promotes better Flexibilityefficiently excited at 405 nm or 488 nm, maximizing violet laser use Minimal single-laser compensationnarrow emission spectra allow for minimal compensation when
using a single excitation source Learn more about Qdot nanocrystals and their applications in flow cytometry and find Qdot nanocrystal conjugates of antibodies that recognize human CD markers at lifetechnologies.com/qdotinflow and find out more about other fluorescent tools that are compatible with your instruments violet laser at lifetechnologies.com/violettools. reproducibility the number of detectable parameters
1. Qdot 525 conjugate excitation 2. Qdot 565 conjugate excitaiton
Extinction coefcient (M-1cm-1)
12,000,000 10,000,000 8,000,000 6,000,000 4,000,000 2,000,000 0 400 500 600 700 Wavelength (nm) 800 900 1,000
3. Qdot 585 conjugate excitation 4. Qdot 605 conjugate excitation 5. Qdot 625 conjugate excitation 6. Qdot 655 conjugate excitation 7. Qdot 705 conjugate excitation 8. Qdot 800 conjugate excitation 1. Qdot 525 conjugate emission 2. Qdot 565 conjugate emission 3. Qdot 585 conjugate emission 4. Qdot 605 conjugate emission 5. Qdot 625 conjugate emission 6. Qdot 655 conjugate emission 7. Qdot 705 conjugate emission 8. Qdot 800 conjugate emission
Extinction coefficients and emission profiles for selected Qdot nanocrystals. Excitation is presented as extinction coefficient; emission is normalized to maximum peak height.
By treating sequentially with the two reagents in the FIX & PERM kits, cells undergo mild fixation and permeabilization that leaves the morphological scatter characteristics intact. This enables researchers to accurately identify previously undetectable intracellular markers, such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, and immunoglobulins. FIX & PERM reagents:
Fixation and permeabilization for flow cytometry samples
Are designed for use with all commercially available flow cytometers Allow the efficient detection of a wide variety of markers and intracellular proteins Help reduce the amount of background staining observed
Find protocols and ordering information online at lifetechnologies.com/fixperm.
105
Cell analysis
We offer a wide variety of Molecular Probes labeling and detection products to aid researchers in producing the best images possible. Some of our most popular productsAlexa Fluor dyelabeled secondary antibodiescan be found in labs across the world and have been foundational tools for fluorescence imaging for decades. We have also developed a number of kits that make it easy for you to directly label your own antibodies for use in fluorescence microscopy and other applicationsfrom the 10-minute labeling offered by Zenon technologies to kits that allow you to label anywhere from 10 g up to 1 mg of antibody. We have developed streptavidin conjugates that deliver moderate signal amplification from your stained samples and Tyramide Signal Amplification (TSA) kits that allow you to detect very low-abundance targets. Learn about all of the Molecular Probes options for immunodetection at lifetechnologies.com/antibodies.
Molecular Probes labeling and detection products
Secondary antibodies and labeling strategies
Immunodetection strategies
Strategy
Use the table below to choose the best immunodetection strategy for your imaging experiment.
Direct detection with labeled antibody Labeled secondary antibody to detect a bound (unlabeled) primary antibody Alexa Fluor 488 Goat Anti-Mouse IgG (H+L) (Cat. No. A11001)
B Fluorophore-conjugated secondary antibodies
Streptavidin or other avidin products to detect biotinylated primary or secondary antibody for moderate amplification Streptavidin, Alexa Fluor 488 conjugate (Cat. No. S32354)
C Biotinylated secondary antibodies
Enzymatic signal amplification for greater amplification
Example product
Alexa Fluor 488 Antibody Labeling Kit (Cat. No. A20181)

A
TSA Kit #22, with HRPstreptavidin and Alexa Fluor 488 tyramide (Cat. No. T20932)
D Enzyme ampli cation
Antigen Primary Ab
Directly conjugated primary antibodies
Secondary Ab Biotin streptavidin complex Enzyme Fluorophores (14 dyes)
(~2 8 dyes/target)
(~1020 dyes/target)
(~2040 dyes/target)
(~120200 dyes/target)
Process time Sensitivity
When to use
Best for abundant targetin samples where secondary detection may generate background. Learn more about antibody labeling kits at lifetechnologies.com/ ablabeling
Best for standard, everyday immunofluorescence. Find labeled secondary antibodies at lifetechnologies.com/ antibodies
Best when used with biotinylated secondary for moderate amplification, or with a biotinylated primary antibody. Learn more at lifetechnologies.com/ streptavidinconjugates
Enzyme-driven signal amplification is best for detecting low-abundance targets. Learn more at lifetechnologies.com/ tsadetection
106
Cell analysis
Create your own labeled antibodies
We offer a number of Molecular Probes labeling kits for the direct attachment of intensely fluorescent Alexa Fluor dyes, R-phycoerythrin (R-PE), or even biotin to less than 1 g to up to 1 mg of IgG antibody. Although the signals from directly labeled antibodies may not be as bright as those observed when using secondary antibodies, their use eliminates the noise commonly observed when secondary antibodies bind nonspecifically to the sample. Also, directly labeled antibodies allow you to use more than one same-species antibody in a single staining experiment. The table below will help you choose from our comprehensive selection of innovative, time-tested, and competitively priced antibody and protein labeling kits that feature streamlined protocols for labeling and conjugate purification. Go to lifetechologies.com/ablabeling to find more information on all of these antibody labeling options.
Amount of IgG <120 g
Product Zenon IgG Labeling Kits APEX Antibody Labeling Kits Small-Scale Protein Labeling Kits
Notes Antibodies ready to use in 10 min Isotype-specific labeling Compatible with stabilizing proteins Antibodies ready to use in 2 hr (~15 min hands-on time) Compatible with stabilizing proteins Antibodies ready to use in 2 hr (~30 min hands-on time) Optimized for 10150 kDa proteins, including IgG antibodies (~150 kDa) Stabilizing proteins must be removed from sample before labeling
Label attachment* Noncovalent
Available fluorophores and labels** Organic dyes, phycobiliproteins Organic dyes
Applications FC, ICC
1020 g
Covalent
FC, ICC, IHC
10100 g
Covalent
Organic dyes
FC, ICC, IHC
100 g
Monoclonal Antibody Labeling Kits
Antibodies ready to use in 90 min (~15 min hands-on time) Optimized for both monoclonal and polyclonal IgG antibodies Stabilizing proteins must be removed from sample before labeling
Covalent
Organic dyes
FC, ICC, IHC
0.53 mg
SAIVI Antibody Labeling Kits
Antibodies ready to use in 75 min (~10 min hands-on time) Includes degree-of-labeling regulator Uses no organic solvents and produces azide-free conjugates
Covalent
Organic dyes
ICC, IHC, in vivo imaging, FC
1 mg
Large-Scale Protein Labeling Kits
Antibodies ready to use in 2 hr (~30 min hands-on time) Designed to label monoclonal and polyclonal IgG antibodies Stabilizing proteins must be removed from sample before labeling
Covalent
Organic dyes
FC, ICC, IHC
FC = flow cytometry; ICC = immunocytochemistry; IHC = immunohistochemistry. * For additional information on covalent vs. noncovalent labeling, see note and diagram below. ** Organic dyes include fluorophores like Alexa Fluor dyes as well as fluorescein and tetramethylrhodamine. Phycobiliproteins include R-PE, APC, and their tandems (e.g., Alexa Fluor 680R-phycoerythrin (R-PE)).
Covalent vs. noncovalent antibody labeling Covalent attachment of a label to an antibody generates a stable conjugateone that lasts several months to years. Labels that are attached noncovalently can dissociate over timethese conjugates last from several hours up to days.
Noncovalent Zenon antibody labeling (A) vs. covalent microscale, monoclonal, and protein labeling (B).
107
Cell analysis
Effective labeling of small amounts of IgG
Many IgG antibodies are available only in small quantities and are often packaged with stabilizing proteins, such as bovine serum albumin (BSA), or are present in whole serum, ascites, or complete tissue culture media. These contaminants can interfere with the labeling reaction. Both the Zenon and APEX antibody labeling kits provide a convenient means to label small amounts of IgG antibodies <20 g with methods that are compatible with these contaminants. Zenon Antibody Labeling Kits Zenon labeling technology provides a versatile, easy-to-use system for labeling mouse IgG1, IgG2a, IgG2b, and rabbit IgG antibodies with our premier Molecular Probes dyes as well as other fluorophore, biotin, photo-proteins, and enzyme labels. This new technology offers several advantages over direct chemical labeling, Including:
Target-specic IgG antibody
Zenon labeling reagent (labeled Fab fragment) Incubate
Labeled Fab fragments bind to the IgG
Labeled IgG ready in only 10 min
Mix with nonspecic IgG, which complexes with unbound Fab fragments
Speedentire labeling procedure typically takes 10 minutes Efficiencylabel nearly 100% of the primary antibody Economylabel submicrogram amounts of antibody Simplicityno pre- or post-labeling purification of the anti Flexibilityeasily use multiple primary antibodies in a single
experiment Learn more at lifetechnologies.com/zenon. APEX Antibody Labeling Kits The new APEX Antibody Labeling Kits provide a convenient means to directly attach a fluorophore to very small amounts of IgG antibody (1020 g), while allowing you to easily remove contaminants without losing antibody. APEX Antibody Labeling Kits utilize a solid-phase labeling technique that captures the IgG antibody on the resin inside the APEX antibody labeling tip. This enables covalent labeling of antibodies that are supplied in solutions containing stabilizing proteins (i.e., BSA) or other contaminants, which can interfere with the amine-reactive labeling reagents that are used to attach the fluorophore to the antibody. With the APEX Antibody Labeling Kit technology, contaminants are simply eluted through the tip. After applying the amine-reactive label, the fluorescent IgG conjugate is ready for use in an imaging (see figure) or flow cytometry application in as little as 2 hours with very little hands-on time. Learn more at lifetechnologies.com/apex. body is required
The Zenon labeling scheme. An unlabeled IgG is incubated with the Zenon labeling reagent, which contains a fluorophore-labeled Fab fragment. The labeled Fab fragment binds to the Fc portion of the IgG antibody, and the excess Fab fragment is bound by the addition of a nonspecific IgG. The addition of nonspecific IgG prevents cross-labeling of the Fab fragment in experiments where multiple primary antibodies of the same type are present. Note that the Fab fragment used for labeling does not need to be coupled to a fluorophore, but could instead be coupled to an enzyme or to biotin.
Cap
Resin
Tip sheath
APEX Antibody Labeling Kits allow convenient elution of contaminants through the filter tip.
108
Cell analysis
Labeled secondary antibodies
Labeled secondary antibodies form stable and specific complexes with unlabeled primary antibodies, providing the foundation for most immunofluorescence microscopy protocols. Molecular Probes extensive selection of labeled secondary antibodies includes antibodies labeled with our superior Alexa Fluor dyes, phycobiliproteins, Alexa Fluor dyephycobiliprotein tandem fluorophores, Qdot nanocrystals, biotin, and enzyme labels (horseradish peroxidase and alkaline phosphatase). We also offer antibodies with different immunoreactivities, essential to avoid confounding cross-reactivity when performing simultaneous secondary immunodetection of two or more targets. We also offer F(ab)2 antibodies, which eliminate the Fc receptor interactions that can be observed when using whole IgG.
Secondary antibody selection guide.

Antibody type Adsorbed against When to use Standard whole IgG Human IgG and human serum Best for standard, single-color immunofluorescence when cross-reactivity is not a concern Highly cross-adsorbed whole IgG Bovine IgG, goat IgG, rabbit IgG, rat IgG, human IgG, and human serum Best for multicolor immunofluorescence when cross-reactivity may be a concern F(ab)2 fragment Human IgG and human serum Best for use in immunofluorescence when Fc interaction observed with whole antibody interferes with results
We offer an extensive selection of labeled antibodies and F(ab)2 fragmentsideal for flow cytometry, fluorescence imaging, western blot analysis and more. Use our online Secondary Antibody Selection Tool to explore a variety of secondary antibodies conjugated to Alexa Fluor dyes, HRP, AP, Qdot nanocrystals, biotin, and more. You can specify target IgG class (and form), host, species reactivity, and conjugate type to narrow your results. Find your secondary antibody now at lifetechnologies.com/antibodies.
Find the labeled antibody youre looking for online
109
Cell analysis
Choosing the right fluorophore
Browse our broad range of fluorescent labels to find the one that best suits your experiment. We offer organic dyes such as the Alexa Fluor dyes, phycofluors and their conjugates (often used in flow cytometry), and Qdot nanocrystals, which exhibit exemplary brightness and photostability. Selecting the right fluorescent label requires matching your application and instrument platform to the properties of the dye. The selection guide below in conjunction with our online SpectraViewer can assist you in selecting the right fluorescence label for your experiment.
Detection reagent Examples
Organic dye Alexa Fluor dyes
Phycofluors
Qdot nanocrystals Qdot 585 Qdot 655
Texas Red dye Fluorescein Photostability Typical analysis platform Applications Brightness Recommended mountant
R-phycoerythrin (R-PE) and conjugates Allophycocyanin (APC) and conjugates
++
FC, FM, HCS ICC, IHC
+
FC ICC
+++
FC, FM ICC, IHC, western blot
++
ProLong Gold antifade reagent, SlowFade Gold antifade reagent, PBS lifetechnologies.com/alexa
+++
Not applicable
+++
Qmount Qdot mounting medium, PBS lifetechnologies.com/qdot
Online resource
lifetechnologies.com/ streptavidinrpe
FC = flow cytometry; FM = fluorescence microscopy; HCS = high-content screening (imaging); ICC = immunocytochemistry; IHC = immunohistochemistry
Image-iT FX Signal Enhancer (Cat. No. I36933) dramatically improves the signal-to-noise ratio of immunolabeled cells and tissues, allowing you to clearly visualize targets that would normally be indistinguishable. Background staining typically seen with fluorescent dyes is largely eliminated when ImageiT FX Signal Enhancer is applied to fixed and permeabilized cells (see figure).
Improve signal-to-noise ratio with Image-iT FX Signal Enhancer
Increased label specificity and resolution provided by Image-iT FX Signal Enhancer. Golgi in fixed and permeabilized HeLa cells labeled with antigolgin-97 (A21270) and visualized with green-fluorescent Alexa Fluor 488 goat antimouse IgG (A11001). Actin was stained with red-fluorescent Alexa Fluor 594 phalloidin (A12381); nuclei were stained with blue-fluorescent DAPI (D3571). Cells in the right panel were treated with Image-iT FX Signal Enhancer (I36933), which eliminates nonspecific dye binding.
110
Cell analysis
BackDrop Background Suppressor
BackDrop Background Suppressor (Cat. No. R37603) is a novel reagent from Molecular Probes designed to effectively suppress background fluorescence in live-cell imaging experiments. BackDrop Background Suppressor is a direct addition product that comes in ultraconvenient dropper bottles as a six pack with two vials each of blue, green, and red background suppressors.
Works with fluorescent dyes and proteins Greatly reduced background fluorescence in live-cell imaging experiments Higher signal-to-noise, improved sensitivity, and better results Convenience of direct addition to cells from dropper bottle, no need to remove medium Flexibility to modulate blue, green, or red background signal
Image-iT Fixation/Permeabilization Kit (Cat. No. R37602) is a convenient-to-use fixation/permeabilization kit that contains all the necessary reagents to prepare your cells for antibody staining and imaging. The highquality components help preserve cell morphology and reduce background staining and come in a practical box with single-use vials and easy-to-follow protocols.
Image-iT Fixation/Permeabilization Kit
A complete fixation/permeabilization kit with ready-to-use components and room temperature storage Optimized for secondary antibody labeling, fixed-cell dye staining, and GFP applications Convenient storage box, 1X ready-to-use components with 18-month room temperature stability, and Compatible with analysis of most cellular antigens Reduced background staining
protocol on the inner lid
Easily accessible online, the Fluorescence SpectraViewer displays the spectral features of one or more fluorophores on a single graph and allows you to overlay representations of your instruments excitation source, excitation filters, and emission filters as well. This tool simplifies the process of choosing fluorophores that are compatible with each other in a multicolor experiment and that will also work with your instruments optics and filters. Find it online at lifetechnologies.com/spectraviewer.
111
Protein analysis
Protein analysis
We have the products to help meet your protein research needs whether you are trying to express proteins in prokaryotic or mammalian cells, or when you need to verify that the protein you made is the right target. Novex, Ambion, and Invitrogen brands provide the trusted, reliable, and scalable solutions for your protein research needs. The chart below highlights the most popular products and new technologies in this chapter. Follow the Web links provided in this chapter for a complete listing of all our products and services. If you have any questions about which of our solutions are right for you, please contact Life Technologies technical support by phone, email, Facebook at facebook.com/novexprotein, facebook.com/ambion, or facebook.com/invitrogen, or Twitter at twitter.com/everydayprotein.
Section
Protein expression systems

Champion pET Expression System, page 119 Insect expression, page 117 PichiaPink Yeast Expression System, page 118 GeneArt Algae Engineering Kit, page 120 GeneArt Gene Synthesis, page 121
Transfection
Lipofectamine LTX Reagent, page 124 Lipofectamine RNAiMAX Reagent, page 125 Lipofectamine 2000 Reagent, page 125 FreeStyle MAX Transfection Reagent, page 125
RNAi
Silencer Select siRNA, page 126127 Stealth Select RNAi siRNA, page 126127 mirVana miRNA Mimics and Inhibitors, page 127128
Protein analysis
Key technologies
Everyday essentials
Lipofectamine Reagents, page 125 Invivofectamine 2.0 Reagent, page 125
siRNA and miRNA, page 126128 Silencer siRNA, page 126 Ambion Pre-miR Precursors and Anti-miR Inhibitors, page 127128
Neon Transfection System, page 124
Instruments
112
Protein analysis
Immunocytochemistry of mouse fibroblasts cells labeled with AKT [pS473] ABfinity Recombinant Rabbit Monoclonal Antibody. See page 147 for more on Novex primary antibodies.
Protein sample prep

Dynabeads immunoprecipitation, page 129 Dynabeads organelle isolation kits, page 132 PARIS Kit for isolation of total RNA and protein, page 133
Protein analysis
Bolt gels for western blotting, page 136138 NuPAGE gels, page 136, 138 Sharp and SeeBlue Plus 2 Pre-Stained Standards, page 141 SYPRO Ruby Protein Gel Stain, page 142 SilverQuest Protein Gel Stain, page 142 iBlot Western Detection Kits, page 145 ABfinity rabbit monoclonal antibodies, page 147
Protein detection
Novex multiplex assays, page 153154 Novex multiplex magnetic assays, page 154155 Novex chemiluminescent ELISAs, page 151
Protein analysis
Protein A/G purification, page 130 Novex proteases, page 131
Novex Tris-glycine gels, page 136 Novex primary antibodies for western blotting, page 146
Novex Antibody Pair kits, page 150 Novex ELISA kits, page 150152
Specialty resins for large and small protein biomolecules, viruses, and antibodies, page 131 Novex cell lysis reagents, page 133
SeeBlue Pre-Stained Standard, page 141 SimplyBlue SafeStain, page 142
POROS Analytical Chromotography, page 131 ZOOM IEF Fractionator, page 132
Qubit 2.0 Fluorometer, page 134 XCell SureLock protein electrophoresis cells, page 139
Luminex MAGPIX System, page 155 Luminex 200 System, page 155 FLEXMAP 3D High-Throughput System, page 155
iBlot 7-minute blotting system, page 143 BenchPro 4100 Western Processing System, page 144
113
Protein analysis
Protein expression systems

Recombinant protein expression technology enables analysis of gene regulation and protein structure and function. Utilization of recombinant protein expression varies widelyfrom investigation of function in vivo to large-scale production for structural studies and biotherapeutics drug discovery. Using the right expression system for your specific application is key to your success. Protein solubility, functionality, speed, and yield are often the most important factors to consider when choosing an expression system. With the wide variety of expression systems available from Life Technologies, youre sure to find one that meets your needs. The following table summarizes some of the characteristics of the most popular expression hosts. Several expression systems from Life Technologies are detailed on the following pages. For a complete list of expression systems, visit lifetechnologies.com/proteinexpression.
Expression hosts and their applications

Host organism Cell-free prokaryotic Most common applications Rapid expression screening Toxic proteins Incorporation of unnatural labels or amino acids Functional assays Protein interactions Advantages Rapid expression directly from plasmid Open systemeasily add components to enhance solubility/functionality Simple format Scalable Compatible with Gateway cloning Prokaryotic Structural analysis Antibody generation Functional assays Protein interactions Yeast Structural analysis Antibody generation Functional assays Protein interactions Insect Functional assays Structural analysis Antibody generation Scalable Low cost Simple culture conditions Compatible with Gateway cloning Eukaryotic protein processing Scalable up to fermentation (grams/liter) Simple media requirements Posttranslational modifications similar to mammalian systems Greater yield than mammalian systems Compatible with Gateway cloning Mammalian Functional assays Protein interactions Antibody generation Highest level of correct posttranslational modifications Highest probability of obtaining fully functional human proteins Compatible with Gateway cloning Multimilligram/liter yields only possible in suspension cultures More demanding culture conditions
Higher Probability of obtaining functional protein
Challenges Large-scale expression (3 mg)
Overall cost and time Lower
Protein expression
Protein solubility Minimal posttranslational modifications May be difficult to express functional mammalian proteins Fermentation required for very high yields Growth conditions may require optimization More demanding culture conditions
114
Protein analysis
Protein expression custom services
Let the experts help you express your protein at the quality and quantity you need Expression optimization Protein production and scale-up Protein purification Life Technologies offers a variety of protein expression, optimization and purification services tailored to meet your individual needs. Our experienced, highly trained scientists can work with you to select the expression systems that best suit your applications and then perform all the steps necessary for obtaining high-quality preparations of your protein of interest. For additional information or price quotations, contact us via phone at 800 955 6288 x45682 (in North America), via email at custom.services@lifetech.com, or visit us online at lifetechnologies.com/customservices.
Selecting a mammalian expression system

1. How much protein do you need? Nanograms to micrograms Milligrams to grams
There are many options in selecting a mammalian expression system matched to your specific needs. Ask yourself the three following questions, then use the table below to match the best system to meet your needs.
Protein yields are highly variable and depend largely on the specific protein to be expressed. In general, small amounts of protein can be readily generated using transient transfection of plasmid DNA into a wide variety of cells. It is very important to achieve the highest transfection efficiency possible using effective transfection reagents such as Lipofectamine LTX or Lipofectamine 2000 reagents. To produce larger amounts of protein (milligrams to grams) requires more cells. Thus, stable cell lines are often employed to produce a large, selected population of cells that is stably expressing your protein of interest. Alternatively, large-scale transient transfection of suspension-adapted cells such as HEK 293 or CHO cells can generate large amounts of protein in a much shorter time period than can be achieved with stable cells. See the following section to understand the choices for producing large amounts of protein. 2. How quickly do you need to obtain your protein? Scalable transient expression delivers high quantities of protein in a short period of time Stable cell lines produce the highest amounts of protein but require a longer time to set up Expression by transient transfection results in high levels of expression for a few days to a week. Thus its ideal for rapid protein production and quick data generation. Transient expression systems such as the FreeStyle 293, FreeStyle MAX CHO, and Expi293 Expression System use suspension-adapted 293 or CHO cells at culture scales from 1 mL to 100 L to generate large amounts of protein. The new Expi293 Expression System has been demonstrated to produce up to 1 gram of protein per liter of cell culture. To produce a cell line that can be used over a long experimental time course or over many experiments, it is necessary to generate a stable cell line in which your expression construct is integrated into the host genome. A selection agent, added to the medium, is used to select the cells that have integrated the construct. If you want to make cell lines with targeted integration into an expression hotspot, we recommend the Jump-In System. For randomly integrated stable cell lines, any pcDNA vector delivered by standard cationic transfection reagent, or the Neon Transfection System, or the ViraPower Lentiviral Expression System may be used.
Protein expression
115
Protein analysis
3. Is it important for you to control when expression begins? Expression systems with constitutive promoters do not allow you to control expression Expression systems with inducible promoters require you to add an inducer to begin expression If you are working with a nontoxic gene and the timing of expression is not important, choose a constitutively expressing promoter. An inducible promoter allows you to control the timing of gene expression. In the absence of an inducer, your gene is not expressed. Add inducer to turn on expression. This option is ideal for expressing toxic proteins. Choose from the T-REx Inducible Expression System or the Flp-In T-REx System. For more information on inducible promoters, please visit lifetechnologies.com/mammalianexpression.
Application Constitutive expression (CMV promoter) Transient expression Scalable, transient expression
System pcDNA vectors pcDNA vectors and cationic transfection reagents Adenoviral Expression Systems FreeStyle 293 Expression System FreeStyle MAX CHO Expression System Expi293 Expression system
Key features Constitutive CMV expression with your choice of several epitope tags and selection markers High-level gene expression in any dividing or nondividing mammalian cell type Scalable suspension culture transfections from 1 mL to 100 L Obtain protein in several days to 1 week Production of up to 1 gram of protein per liter of culture
Protein expression
Stable cell lines, targeted integration
Jump-In Fast Gateway System Jump-In TI-Gateway System Flp-In T-REx System
Rapid generation of stable cell lines with integration into expression hotspots Generation of stable cell lines with integration into preselected genomic sites Regulated expression from CMV promoter Rapid generation of regulated stable expression cell lines Regulated expression in any dividing or nondividing mammalian cell type
Inducible expression
T-REx System, Flp-In T-REx System ViraPower Lentiviral T-Rex System
Find out more about mammalian expression systems at lifetechnologies.com/mammalianexpression.
116
Protein analysis
Insect expression systems

High yields, more functional proteins
Expression in insect cells offers significant advantages, including high expression levels, ease of scale-up, production of proteins with proper posttranslational modifications, and simplified cell growth. Insect cells do not require CO2 for growth and can be readily adapted to high-density suspension culture for large-scale expression. Many of the posttranslational modification pathways present in mammalian systems are also utilized in insect cells, allowing the production of recombinant protein that is antigenically, immunogenically, and functionally similar to the native mammalian protein. Baculovirus expression systems are powerful and versatile systems for high-level recombinant protein expression in insect cells. Expression levels up to 500 mg/L have been reported using the baculovirus expression system.
The BaculoDirect Baculovirus Expression System is the fastest and easiest method available for
generating recombinant baculovirus. The BaculoDirect Linear DNA includes attR sites for rapid and efficient recombinational cloning with a Gateway entry clone. The resulting recombinant DNA is taken directly from the LR reaction mix and used to transfect insect cells, which helps to save a significant amount of time. Purified virus can typically be isolated within one week. The reduction of hands-on time makes BaculoDirect ideal for high-throughput expression. The Bac-to-Bac Baculovirus Expression System uses a unique bacmid shuttle vector that recombines by site-specific transposition and generates an expression bacmid in bacterial cells. The expression bacmid is then transfected into insect cells to generate recombinant baculovirus. The Bac-N-Blue Baculovirus Expression System has been used for over a decade to produce high levels of recombinant proteins. The Drosophila Expression System (DES) uses the well-characterized Drosophila Schneider S2 cells and simple expression vectors to allow stable or transient expression of recombinant proteins.
Protein expression
Selection guide for insect expression systems.

Fusion partner System BaculoDirect Host Sf9, Sf21, or High Five Sf9, Sf21, or High Five Sf9, Sf21, or High Five S2 cells Honeybee melittin Secretion signal Position N-term C-term Purif. 6xHis 6xHis Epitope V5 V5 Promoter Polyhedrin Expression/ inducer Infection Advantage Fast and easy method for generation of recombinant baculovirus; ideal for high throughput Rapid baculovirus production; easy blue/ white selection of recombinant colonies Classic and trusted expression system for high-level recombinant protein production Easy-to-use stable system, constitutive or inducible expression; uses simple plasmids for expression; extremely high integration of transfected plasmids eliminates need to select and screen for expression from clonal cell lines
Bac-to-Bac or Bac-to-Bac HBM
GST N-term
6xHis
pFastBacHT pDEST10
Polyhedrin or p10
Infection production
Bac-N-Blue
Honeybee melittin
C-term
6xHis
Xpress V5
Polyhedrin
Infection
DES
BIP
C-term
6xHis
V5
MT or Ac5
CuSO4 or constitutive
Find out more about insect expression systems at lifetechnologies.com/insectexpression.
117
Protein analysis
PichiaPink Yeast Expression System
Large-scale production of secreted recombinant proteins

Screen Pichia transformants rapidly by adenine auxotrophy and color selection Reduce protein degradation with protease-deficient Pichia pastoris strains Optimize protein secretion with one of eight secretion signal sequences
The PichiaPink System is a eukaryotic protein expression system based on the eukaryote Pichia pastoris, which can be used for high-level (grams per liter) and largescale (>1,000 L) production of secreted recombinant proteins. This system contains both low- and high-copy plasmid backbones, eight secretion signal sequences, and four yeast strains to help you optimize for the highest possible yield of recombinant protein. P. pastoris carries out many of the same posttranslational modifications that occur in mammalian cells, which means that the likelihood of producing a functional recombinant protein from this expression system is very high. Additional advantages of P. pastoris include rapid growth, a well-defined genetic background, simple media formulations, and simple handling techniquesall of which make it the ideal host for producing large quantities of protein in a short period of time. The PichiaPink System provides you with two different kits for secreted expression of your recombinant protein of interest:
Use color selection to find your transformed Pichia Pink colonies. Transformed Pichia appear as white colonies, while untransformed Pichia appear as redpink colonies.
Protein expression
The PichiaPink Secretion Optimization Kit enables you to screen multiple signal
sequences with your gene of interest in both low- and high-copy vectors (pPink-LC and pPink-HC, respectively) for optimal expression and secretion of your recombinant protein. pPink-LC and pPink-HC vectors are also available separately as the PichiaPink Vector Kit. The PichiaPink Secreted Protein Kit allows you to clone your gene interest in frame with the Saccharomyces cerevisiae -mating factor presequence using the pPink-HC plasmid for secreted expression of your recombinant protein. pPink-HC is also available separately as the PichiaPink Secreted Protein Vector Kit. PichiaPink Secretion Optimization Kit contents: PichiaPink Secreted Protein Kit contents:
The PichiaPink Vector kit containing

a high- and low-copy plasmid The PichiaPink Secretion Signal set containing 8secretion signals The PichiaPink Expression Strain set containing 4glycerol strains The PichiaPink Media Kit containing 4 basic media
The PichiaPink Secreted Protein

Vector kit containing the highcopy plasmid with the S. cerevisiae -mating factor presequence to yield secreted protein expression The PichiaPink Expression Strain set containing 4glycerol strains The PichiaPink Media Kit containing 4 basic media
Find out more about yeast expression systems at lifetechnologies.com/yeastexpression.
118
Protein analysis
Prokaryotic expression systems
Easy-to-use recombinant protein expression systems

Due to their ease of use, relatively high yields, and simple scalability, T7-regulated E. coli expression systems are often chosen for generating recombinant protein. Selecting the right system for your specific application is the key to your success. With the wide variety of T7 expression systems available from Life Technologies, youre sure to find one that meets your needs. The table below summarizes some of the most popular T7 expression systems. For a complete list, go to lifetechnologies.com/proteinexpression.
Champion pET Expression Systems offer the high-level expression

Protein of interest constitutes >50% of total cellular protein 5-minute TOPO Cloning with >95% efficiency No ligase, post-PCR procedures, or restriction enzymes required
The Champion pET Expression System uses optimized vectors and a BL21 Star E. coli expression strain to produce protein yields up to 10-fold greater than any other expression system. The system takes advantage of the high activity and specificity of the bacteriophage T7 RNA polymerase for high-level transcription of the gene of interest. The lac operator located in the promoter region provides tighter regulation than traditional T7-based vectors, improving plasmid stability and cell viability. BL21 Star E. coli are genetically engineered to reduce transcript degradation, resulting in significantly improved mRNA stability and increased protein production. Cloning into a Champion pET Expression vector is easy, fast, and efficient. Available with Directional TOPO or Gateway Cloning technologies, you can typically generate your clone in one day. In addition, the Champion pET vectors offer simplified protein purification, protein detection, enhanced solubility, and native protein expression.
Protein expression
Selection guide for our most popular T7 expression systems.

System Champion pET302/NT-His and pET303/CT-His Vector Kit Champion pET300/NT-DEST and pET301/CT-DEST Gateway Vector Kit Champion pET Expression System Champion pET SUMO Expression System Champion pET Expression System Champion pET BioEase Expression System T7 Expression System pBAD Expression System trc Expression System Promoter T7 lac T7 lac T7 lac T7 lac T7 lac T7 lac T7 araBAD trc Advantage Higher expression levels than those obtained from other pET vector suppliers Convenience of Gateway cloning and choice of N-term or C-term 6xHis tag Highest level of protein production Highest level expression of proteins with enhanced solubility Rapid, visual screening of protein expression without western blotting with Lumio technology Easy, efficient expression of biotinylated proteins High-level expression of nontoxic proteins Tightly regulated expression for efficient expression of difficult-toexpress proteins Enhanced expression of eukaryotic protein
Find out more about prokaryotic expression systems at lifetechnologies.com/bacterialexpression.
119
Protein analysis
GeneArt Algae Engineering Kits

Genetic modification and expression systems for photosynthetic microalgae
GeneArt Algae Engineering Kits for Chlamydomonas reinhardtii and Synechococcus elongatus are the first commercially available genetic modification and expression systems for photosynthetic microalgae. These kits are designed for rapid scale-up and production and consistent, defined quality.
Algal cells arrive ready to resuscitate, grow, and transform or store at 80C until ready to use Every cell lot is manufactured using a standardized manufacturing protocol, so every experiment starts Optimized media, vectors, and protocols help ensure robust selection and expression
with consistent materials
Get transformed cells more quickly
GeneArt Algae Engineering Kits save time required for strain optimization and transitioning to downstream applications such as bioproduction.
Protein expression
Cells are typically ready for transformation in less than 5 days GeneArt vectors facilitate rapid directional cloning of synthetic genes or PCR products Gibco media are provided at 1X concentration, eliminating time-consuming media preparation
Dont wonder about the condition and characteristics of your cultures and expression clones. With GeneArt Algae Engineering Kits, you have access to a quality-controlled supply of algae cells and cloning tools.
Take control of the quality of your reagents
Kits are shipped and stored at 80C Algal cell stocks are viable, pure, and standardized GeneArt vectors are fully characterized and optimized for engineering and expression in each strain
Find out more about algae expression systems at lifetechnologies.com/algaeexpression.
Thaw and resuscitate algae (24 days)
50 45
40
35
30
25
Grow algae in light for 24 days to OD >0.6

750
Transform algae and test for expression (56 days)
20
15
10 7.5 5
Construct and verify the expression clone (3 days)
Amplify the gene of interest, insert into vector, and conrm
GeneArt Algae Engineering Kits offer quality-controlled components and significant process efficiencies for both research and bioproduc tion environments. Following the optimized workflow, you can typically see transformed cells in less than 10 days.
120
Protein analysis
GeneArt gene synthesis service
Let us synthesize your DNA constructs for improved or engineered protein expression
Gene synthesis has become the most cost-effective, time- and resource-saving method for obtaining nearly any desired DNA construct, producing expression clones that outperform those produced using conventional molecular biology techniques in expression performance, stability, and quality. GeneArt gene synthesis tools go beyond traditional synthesis and enable you to:
Improve protein expression with proprietary GeneOptimizer technology Gain access to hard-to-clone constructs, long, complex DNAs, and customized vectors Create unlimited numbers of mutants for screening experiments Overcome RNAi inactivation Engineer proteins to improve enzymes and humanize and/or increase binding affinities of antibodies
Find out more about gene synthesis at lifetechnologies.com/genesynthesis.
G
A
A
G
Protein expression
www
5 5 5 5 5 5 5 5 5 5 5 5 5 5
A C G C A
Day 1 Ordering until 3:00 pm (CET) Oligosynthesis over night Day 2 Gene Assembler Day 3 Cloning Day 4 Sequencing & quality control Day 5 Ready for shipment
GeneArt SuperSPEED production schedule.
Our proprietary GeneOptimizer algorithm offers proven increases in protein yield through sequence optimization. By evaluating many important expression parameters in parallel, GeneOptimizer technology generates up to 500,000 variants of your target sequence in an evolutionary approach and selects the best match for your specific requirements. Sequence optimization using the GeneOptimizer software process is included as an optional step with all GeneArt gene synthesis services. Select GeneOptimizer technology when you need:
GeneArt GeneOptimizer sequence optimization
Removal of introns Knockout of cryptic splice sites and RNA destabilizing sequence elements Increased RNA stability Adaptation of codon usage Extensive mutagenesis Flexible combination of functional domains Introduction of restriction sites Epitope shuffling Consideration of immunomodulatory CpG motifs
Find out more about gene optimization at lifetechnologies.com/geneoptimization.
121
Protein analysis
ty pe Op tim iz ed d Op tim iz e ty pe ild W
M oc k
ng Pr55/g total protein
3,5 3 2,5 2 1,5 1 0,5 0
Capture assay
M oc k
ild
Northern
Remove sequence repeats

PABP
PABP
Adjust codon usage
Optimize GC content
gag
PABP
AAAAAA
AAAAAA
Avoid RNA secondary structure
Western
Eliminate killer motifs
Remove splice sites
Beta-actin
Optimized gene
Sequence optimization using the GeneArt GeneOptimizer process significantly increases expression. Reference: J Virol 74:10822 (2000).
Proprietary GeneOptimizer algorithm determines optimal gene sequence for your experiments.
Selection guide for protein expression clones and expression optimization services.
Protein expression
Order now, express in a week 100% sequence-verified clone available in less than a week Human Ultimate ORF Clones https://orf.invitrogen.com/ cgi-bin/ORF_Browser
Configure your projectprotein expression with no gene design constraints GeneArt gene synthesis 817003DE (0.53 kb, online ordering) 817000DE (<0.5 kb, online ordering) Full service that delivers validated genes with no limitation in sequence design 615 days to receive ready-to-use clones Up to 20 kb NA Synthetic wild type gene 5 g lyophilized DNA cloned in GeneArt standard vector Quality assurance documentation, including sequence verification
Optimize your expression improved protein expression levels through gene optimization GeneArt GeneOptimizer sequence optimization 817003DE (0.53 kb, online ordering) 817000DE (<0.5 kb, online ordering) Full service that delivers validated genes, with improved expression yields and no limitation in sequence design 615 days to receive ready-to-use clones Up to 20 kb NA Optimized gene 5 g lyophilized DNA cloned in GeneArt standard vector Quality assurance documentation including sequence verification Any vector Up to 100-fold Yes
Product name Cat. No.
Key features
Fastest way to obtain a sequence verified clone
Time to delivery Gene Size Number of fragments What is delivered
1 week One gene 1 Glycerol stock of uORF entry clone ORF card data available online
Choice of expression vectors Free web-based tool for search and design
Any Gateway destination vector Any vector NA Yes Yes
Improvement of expression yields NA
122
Protein analysis
Overview of transfection
Life Technologies reagent-mediated and electroporation-based transfection products are the most cited and trusted in the industry. Regardless of your cell type, throughput requirements, or budget, we have the right solution for you.
Fits your experimentwhether youre transfecting plasmid DNA, siRNA, mRNA, miRNA, RNAi plasmids, or protein expression Easy to useall of our products come with clear and simple protocols for maximum convenience and ease of use Works with your cellsour transfection systems and reagents were designed for efficient transfection of a broad range of cell Used by scientists worldwideLife Technologies is the most cited and trusted supplier of transfection reagents, with over
48,000 citations for the Lipofectamine brand alone types, including adherent, suspension, and difficult-to-transfect cells constructs, we have the right product for you
The table below outlines our most popular transfection products. If you go to lifetechnologies.com/transfection, you can find information about our entire line of transfection products and youll also have access to:
Selection guides and performance data for choosing the correct transfection product Product tables sorted by sample type and transfection method Cell typespecific transfection protocols for more than 120 cell lines FAQs and information on selection antibiotics to help you prepare for your experiment
Selection guide for highlighted transfection reagents and systems. Find this interactive table online at lifetechnologies.com/lipofectamine.
Value reagent for common cell types Most cited and versatile reagent for a wide range of cell types Most efficient reagent for plasmid delivery; potent and low cytotoxicity Lipofectamine LTX Most efficient reagent for siRNA/ miRNA delivery; efficient gene knockdown Lipofectamine RNAiMAX Cat. No. 13778075 RNA (e.g., siRNA, miRNA) High-efficiency electroporation for all cell lines
Transfection and RNAi
Lipofectamine
Lipofectamine 2000
Neon Transfection System Cat. No. MPK5000S Plasmid DNA RNA (e.g., siRNA, miRNA) Protein RNAi plasmids
Top-selling format Sample type
Cat. No. 18324012 Plasmid DNA
Cat. No. 11668019 Plasmid DNA RNA (e.g., siRNA, miRNA) RNAi plasmids
Cat. No. 15338100 Plasmid DNA RNAi plasmids
Transfection efficiency for common cell types Best for primary and stem cells
Standard
High
Superior
Superior
Maximal
Watch the video (lifetechnologies.com/tfinsights) to learn about the transfection research program and key factors for successful transfections, which resulted in enhanced protocols for the Lipofectamine line of transfection reagents.
123
Protein analysis
Neon Transfection System
cells, primary, and stem cells to-use protocols
Superior transfection efficiency for labs of all sizes, budgets, and needs
Maximum efficiencyup to 90% in many cell types, including difficult-to-transfect Flexibleeasily transfect from 1 x 104 cells to 5 x 106 cells per reaction with easy Simpleeasy to use, with single reagent kit for all cell types Versatileopen system allows electroporation parameters to be optimized freely
Learn more about the advantages of the Neon Transfection System at lifetechnologies.com/neon. B
High transfection efficiency of Jurkat cells with the Neon Tranfection System. Jurkat cells were transfected with an EGFP-encoding vector using the Neon Transfection System and viewed using (A) bright-field microscopy and (B) fluorescence microscopy 24 hr following transfection.
Lipofectamine LTX and PLUS Reagent

viability
Efficient reagent for plasmid delivery and protein expression
Superior performancehigh transfection efficiency with maximal (>90%) cell Easysimple protocol provided for many cell lines Versatilesuperior plasmid delivery into a broad range of cell types including
primary and hard-to-transfect cells Learn more about the advantages of Lipofectamine LTX Reagent at lifetechnologies.com/ltx.
0 ng
500 ng
1,000 ng
Lipofectamine LTX Reagent with PLUS Reagent, efficiently transfects primary neural progenitor cells. Lipofectamine LTX Reagent (1.5 L) was used to transfect murine embryonic primary neural progenitor cells with the indicated quantities of a GFP-expressing plasmid in the presence of PLUS Reagent (2.5 L). GFP expression was analyzed 24 hr post-transfection. Data courtesy of the Beverly Davidson laboratory, University of Iowa.
124
Protein analysis
Lipofectamine RNAiMAX Reagent
Efficient reagent for siRNA or miRNA delivery in vitro, potent gene knockdown
Superior performanceexcellent transfection efficiency and gene knockdown Easysimple protocol, minimal cytotoxicity across a 10-fold concentration range of transfection reagent Versatiledeliver siRNA or miRNA into a broad range of cell types including primary and hard-totransfect cells Learn more about the advantages of Lipofectamine RNAiMAX Reagent at lifetechnologies.com/rnaimax.
Lipofectamine 2000 Reagent
Most cited and versatile reagent for a broad range of cell types, co-transfection
Provenmost cited transfection reagent with exceptional transfection efficiency Easysimple protocol, efficiency in the presence of serumeliminates need to change media following Versatileeffective for both plasmid and siRNA or miRNA delivery and co-transfection
Learn more about the advantages of Lipofectamine 2000 Reagent at lifetechnologies.com/lf2000. transfection in most cases
Lipofectamine Reagent
Value reagent for common cell types
Reliableappropriate for establishing stable cell lines Flexibleproven to work in high-throughput applications Effectiveworks well with PLUS reagent for higher protein expression
FreeStyle MAX Transfection Reagent
Animal origin-free reagent for transient transfection of CHO and HEK 293 cells
High yielddesigned for recombinant protein bioproduction applications Efficientdesigned for the highest expression levels and transfection rates with the lowest cytotoxicity Rapidup to milligram quantities (including all posttranslational modifications) produced in one week
Find out more at lifetechnologies.com/freestyle.
Invivofectamine 2.0 Reagent
Easy-to-use, effective reagent for siRNA or miRNA delivery in vivo
Effectivespecific, effective knockdown (>80%) in mouse liver Simpleeasy-to-use procedure; just mix, equilibrate, and inject Nontoxic and non-immunostimulatory
Find out more at lifetechnologies.com/invivofectamine.
Meet the Inventor: Invivofectamine 2.0 Reagent Watch this video (lifetechnologies.com/inventorinvivo) to learn more about Invivofectamine 2.0 Reagent, the latest innovation for in vivo siRNA delivery from Life Technologies.
125
Protein analysis
Overview of RNAi
Two types of small RNA molecules function in RNAi. The first type comprises synthetic, short interfering RNA (siRNA) molecules that target mRNA cleavage effectively knocking down expression of a gene of interest. The second are microRNA (miRNA) molecules that regulate gene expression by binding to the 3 untranslated regions (UTRs) of target mRNAs to inhibit their function. There are several ways to induce RNAi: synthetic molecules, RNAi vectors, and in vitro dicing. Your choice of tools depends on your model system, the length of time you require knockdown, and many other experimental parameters.
The sections below highlight some of our most popular RNAi products, but you can view even more at lifetechnologies.com/rnai. There youll find:
Guides for choosing the correct RNAi delivery method for your experiment The 160-page RNAi Sourcebook (downloadable PDF) Easy-to-use online ordering tools for custom RNAi synthetics and vectors and custom miRNA inhibitors and precursors Made-to-order and preplated siRNA libraries siRNA positive and negative controls Protocols and guidelines to help you plan and execute your experiment
For vector-based RNAi products, visit lifetechnologies.com/vectorrnai.
% mRNA remaining
Traditional RNAi methods for gene knockdown in mammalian cells involved the use of synthetic RNA duplexes consisting of two unmodified 21-mer oligonucleotides annealed together to form short/small interfering RNAs (siRNAs). Silencer Select siRNA and Stealth RNAi siRNA improve upon these traditional duplexes by using proprietary chemical modifications to ensure better RNAi results. Silencer Select siRNA Highest knockdown, lowest off-target effects 100-fold more potent knockdown Lowest off-target effects via locked nucleic acid (LNA) chemistry 100% guaranteed to silence (>70%) (terms and conditions apply; go to lifetechnologies.com/silencerselect) Stealth Select RNAi siRNA Good knockdown, low off-target effects
siRNA
100 90 80 70 60 50 40 30 20 10 0 % effective 94% 86% 87% 68%
70% Knockdown Silencer siRNA Designs
80% Knockdown Silencer Select siRNA Designs
1 6 11 16 21 26 31 36 41 46 51 56 61 66 71 76 81 86 91 96 101 106 111 116 121 126 131 136 141 146 151
High knockdown Low off-target effects

Silencer siRNA Reliable, affordable siRNA Cost-effective Good target specificity
Silencer Select siRNA design algorithm significantly improves effective siRNA prediction accuracy. The Silencer Select siRNA design algorithm was used to design 155 siRNAs to 40 different targets. These siRNAs were tested side by side with siRNAs designed using the previous algorithm at 5 nM in HeLa cells. mRNA knockdown was measured 48 hr post-transfection via qRT-PCR using TaqMan Gene Expression Assays. Results are expressed as percent of mRNA remaining compared to Silencer Negative Control #1 siRNAtreated cells. The inset shows the percentage of siRNAs that elicited 70% and 80% mRNA knockdown.
126
Protein analysis
Which siRNA is right for you? Find this interactive table online at lifetechnologies.com/siRNA.
Cost-effective siRNA Silencer siRNA Potency Efficacy (>70% knockdown) Target specificity Innate immune response Molecular format Coverage Target species Recommended product 100 nM recommended conc. 2 of 3 siRNA guaranteed Moderate Minimum Unmodified 21 bp duplex with overhangs Coding RNA Human, mouse, rat (use custom tool for other species) Silencer Pre-designed siRNA, 5 nmol (Cat. No. AM16708) Good knockdown, low off-target Stealth Select RNAi siRNA 20 nM recommended conc. 2 of 3 siRNA guaranteed High Minimized through chemical modifications Modified 25 bp duplex with no overhangs Coding RNA Human, mouse, rat (use custom tool for other species) Stealth RNAi siRNA Tube (Cat. No. 1299003) Highest knockdown, lowest off-target Silencer Select siRNA 5 nM recommended conc. 2 of 2 siRNA guaranteed Highest Minimized through chemical modifications LNA-modified 21 bp duplex with overhangs Coding and noncoding RNA Human, mouse, rat (use custom tool for other species) Silencer Select Pre-designed siRNA, 5 nmol (Cat. No. 4392420)
miRNA
MicroRNAs (miRNAs) are short, highly conserved small noncoding RNA molecules naturally occurring in the genomes of plants and animals. miRNAs are 1727 nucleotides long and regulate posttranscriptional mRNA expression, typically by binding to the 3 untranslated region (3-UTR) of the complementary mRNA sequence, resulting in translational repression and gene silencing. Studies have shown that thousands of human protein-coding genes are regulated by miRNAs, indicating that miRNAs are master regulators of many important biological processes. Since the discovery of the first lin-4 miRNA in C. elegans, miRNAs have been identified in diverse organisms through experimental determination or computational prediction. These are managed through the miRBase database, which is a searchable public database of all published miRNA sequences and annotations. mirVana miRNA Mimics and Inhibitors Next-generation miRNA Low off-target effects and high potency 100% coverage of miRBase v 17 In vitro and in vivo use Ambion Pre-miR Precursors and Anti-miR Inhibitors Reliable, affordable miRNA High specificity and efficient inhibition 100% coverage of miRBase v 15
Meet the Inventor: Silencer Select siRNA Susan Magdaleno describes Silencer Select siRNA technology (watch the video at lifetechnologies.com/inventorsirna).
127
Protein analysis
1.40 1.20 1.00
Fold change
FW RV
450 400
mRNA upregulation %
350 300 250 200 150 100 50 0 miR 122
miR122 = miR122 miRNA inhibitor NT = Not transfected Neg1 = Negative Control #1
0.80 0.60 0.40 0.20 0.00 miRNA mimic Neg miRNA mimic Neg miRNA mimic Neg miRNA mimic Neg miRNA mimic Neg miRNA mimic Neg
NT
Neg1
reporter 1
reporter 2
reporter 3
reporter 4
reporter 5
reporter 6
AldoA
miR 122
NT
Neg1
Hfe2
miR 122
NT
Neg1
Slc35a4
miR 122
NT
Neg1
Lass6
mirVana miRNA mimic mature strand is highly potent while star strand is inac tivated. The key advantage of mirVana miRNA mimics is inactivation of the star strand. miRNA mimics, like natural microRNAs have 2 strandsthe mature strand (guide strand) that is functional and used by Ago protein to target mRNAs; and the star or passenger strand, which is nonfunctional and is normally cleaved and expelled from the complex. Most scientists want to analyze one strand of miRNA at a time, and want the other strand to be totally inactive: mirVana miRNA mimics achieve this. For this assay we measured activity from both strands of miRNA mimics. One reporter has a target in forward orientation to measure activity of the mature miRNA strand; another reporter has the target cloned in reverse/complement orientation to test activity of the star strand of the miRNA mimic. For all 6 sequences, activity of the mature strand is high (5- to 10-fold lower than negative control), and activity of the star strand is low or nothing (similar to neg control).
mirVana miRNA Inhibitors effectively suppress miRNA in vivo. miR122 or Negative Control #1 mirVana miRNA inhibitors were complexed with Invivofectamine 2.0 Reagent and delivered to Balb-C mouse liver via tail vein injection on three consecutive days at a dose of 7 mg/kg body weight. Expression levels of four mRNA targets (AldoA, Hfe2, Slc35a4, and Lass6), natural targets of miR122, were measured with TaqMan Assays in the livers of mice injected with miR122 inhibitor, Negative Contol #1 (Neg 1) as well as untreated animals. Significant up-regulation of mRNA was detected in livers of mice treated with miR122, compared to untreated and negative control-treated mice; >3.5-fold for AldoA, >2-fold for Hfe2, >3-fold for Slc35a4, and >4-fold for Lass6. This indicates that mirVana miRNA inhibitors are efficiently delivered to the liver with Invivofectamine 2.0 Reagent and inactivate miR122, leading to up-regulation of genes naturally suppressed by miR122.
Which miRNA mimic or inhibitor is right for you? Find this interactive table online at lifetechnologies.com/miRNA.
Novel design to minimize off-target effects Ambion Pre-miR Precursors Function Content database Model system Species covered Custom design tool miRNA libraries Recommended product Mimic endogenous miRNAs 100% coverage of miRBase v 15* In vitro Human Design your custom miRNA mimics Predesigned or custom libraries Pre-miR miRNA Precursor, 5 nM (Cat. No. AM17100) Next-generation chemistries Chemically modified for for lowest off-target effects good efficacy mirVana miRNA Mimics Mimic endogenous miRNAs 100% coverage of miRBase v 17** In vitro and in vivo Ambion Anti-miR Inhibitors Inhibit endogenous miRNAs 100% coverage of miRBase v 15* In vitro Next-generation chemistries for highest efficacy mirVana miRNA Inhibitors Inhibit endogenous miRNAs 100% coverage of miRBase v 17** In vitro and in vivo Human, mouse, rat (use custom tool for other species) Design your custom miRNA inhibitors Predesigned or custom libraries mirVana miRNA Inhibitor, 5 nM (Cat. No. 4464084)
Human, mouse, rat (use Human custom tool for other species) Design your custom miRNA mimics Predesigned or custom libraries mirVana miRNA Mimic, 5 nM (Cat. No. 4464066) Design your custom miRNA inhibitors Predesigned or custom libraries Anti-miR Inhibitor, 5 nM (Cat. No. AM17000)
* 1,090 distinct human sequences. ** 1,719 distinct human sequences.
Meet the Inventor: mirVana mimics and inhibitors Alexander (Sasha) Vlassov describes mirVana miRNA mimics and inhibitors and their applications for functional analysis (watch the video at lifetechnologies.com/inventormirna).
128
Protein analysis
Protein sample preparation

Life Technologies offers reliable solutions to isolate peptides, proteins and protein complexes directly from your crude sample for further analysis to advance your proteomic workflows. Select the protein sample preparation method for your sample using the table below. Further information on each of the recommended methods can be found in the pages that follow.
Type of sample Cell lysate or tissue homogenate His-tagged recombinants Complex protein mixtures Fusion proteins Bioproduction Whole cells Whole cells Whole cells
Protein type needed Intact proteins and protein complexes Intact, recombinant proteins Intact antibodies, IgG or Ig fragments Intact, untagged proteins Clean, intact, sterile large- or pilot-scale protein for industrial applications Intact protein, separated by isoelectric charge Intact organelle proteins Intact, native protein
Recommended method(s) Dynabeads for immunoprecipitation His-tag purification with affinity resins Antibody purification with protein A/G Protein tag removal with proteases Analytical and production chromatography Protein sample fractionation Organelle isolation kits Cell lysis buffers and protein extraction kits
Dynabeads for immunoprecipitation

Efficient and reliable immunoprecipitation
Dynabeads Protein A or Dynabeads Protein G
Reduce protocol time to ~30 minutes Isolate intact proteins and protein complexes Minimize background caused by nonspecific binding Kits supplied with premade buffers for convenience and
increased consistency
Y
Y
Add sample with antibody
For efficient immunoprecipitation, you need only a Dynabeads Immunoprecipitation Kit, your specific antibody of choice, and a DynaMag magnet. The protocol takes place in a single tube with just a few handling steps. 1. Bind antibodies to the Dynabeads (10 min) 2. Wash once using a magnet (2 min) 3. Mix Dynabeads and sample (e.g., lysate) for immunocapture (10 min) 4. Wash 3 times using a magnet (5 min) 5. Elute the antigenantibody complex in a small volume (2 min), or resuspend directly in SDS/LDS sample buffer, followed by heating 6. Run on a gel
Elute
Y
Add sample with target protein
Isolate pure antibodies
Elute
Mild elution for isolation of native protein
Y Y
Denaturing elution for isolation of denatured target protein
Immunoprecipitation in only 30 minutes. Dynabeads precoupled with protein A or protein G act as a suspendable solid support that can be fixed by the use of a magnet. This allows for simple and efficient antibody capture, followed by immunoprecipitation of your pure target peptides, proteins, protein complexes, or other antigens.
Y Y
Y
Y
129
Protein analysis
Gentle, rapid handling for better results
Immunoprecipitation with Dynabeads is fast and gentle, causing minimal physical stress to target proteins. Rapid kinetics and short incubation times reduce the protocol time typically to only 30 minutes. Dynabeads permit the isolation of labile complexes that might otherwise dissociate or be damaged by proteases during long incubations. Native protein conformation and intact, large protein complexes are preserved. Gentle magnetic handling allows you to work with concentrated protein solutions throughout the procedure. You do not need to worry about losing material from spun-down resins or excess surface during pipetting. Washing and elution are very efficient and help ensure maximal sensitivity, minimal loss of target protein, and no background caused by nonspecific binding.
The protocol can be scaled to match your specific purpose and sample volume requirements. An important advantage of short incubation times and the low nonspecific binding characteristics of Dynabeads products is that there is no need for time-consuming preclearing or dilution.
Easy, flexible, and automatable
Find more information online Protein sample preparation
The table below outlines our most popular immunoprecipitation products. If you go to lifetechnologies.com/ immunoprecipitation, you can find information about our entire line of immunoprecipitation products and youll also have access to:
Product selection guides for immunoprecipitation (IP) and co-IP Yield, reproducibility, and binding specificity data for Dynabeads products Protocols for IP crosslinking and other applications Information to help you choose the right assay format and magnet References that cite the use of Dynabeads for products IP and protein isolation
Dynabeads products for immunoprecipitation. Find the immunoprecipitation product you need using our interactive selec tion guide at youtube.com/immunoprecipitation.
Dynabeads product Protein A (for IP) Protein G (for IP) Co-IP kit for protein complexes His-tag isolation and pulldown Antibody Coupling Kit Bead characteristics Surface:protein A (Cat. No. 100-06D) Surface:protein G (Cat. No. 100-07D) Surface: epoxy (Cat. No. 143-21D) Surface: cobalt NTA chemistry (Cat. No. 101-03D) Surface: epoxy (Cat. No. 143-11D) Binding properties Noncovalent antibody binding Noncovalent antibody binding Covalent antibody binding (antibodywill notelute off beads) Ionic interaction, very low background Covalent binding of antibodies andother desired proteins Type of ligand Manyantibodies Many antibodies Any antibody Binds histidine-tagged proteins Binds any antibody andprotein ligand Coupling >10 min room temp 1040 min room temp 15 min setup (overnight reaction) 5 min room temp 15 min setup (overnight reaction)
130
Protein analysis
His-tag purification with affinity resins
Life Technologies offers affinity resins and purification kits for use with 6xHis-tagged proteins. Purification of histidine-tagged recombinant proteins can be performed with precharged Ni resin (Ni-NTA agarose beads) or with an iminodiacetic acid ligand (ProBond Nickel Chelating Resin). To select your product, visit lifetechnologies.com/histagpurification.
Antibody purification with protein A/G
Life Technologies offers a variety of affinity resins for antibody purification. IgG purification, as well as fractionation of IgG fragments, can be achieved using Protein A Agarose, Protein G Agarose, or Streptavidin Agarose. To learn more or select the product you need, visit lifetechnologies.com/antibodypurification.
Protein tag removal with proteases
Life Technologies offers a wide selection of proteases for efficient removal of affinity tags and cleavage of fusion proteins. Recombinant proteins that enterokinase, or SUMO recognition sequences provide the option for protein tag removal with highly specific and active cleavage enzymes.
Protease Cat. Nos. Enzyme
AcTEV Protease 12575015, 12575023 Tobacco Etch Virus (TEV) with improved specificity, activity, and stability than native TEV protease; contains 6xHis for easy removal Glu-Asn-Leu-Tyr-Phe-Gln-Gly Cleavage of any fusion protein containing the recognition sequence
EKMax Enterokinase E18001, E18002 Recombinant bovine enterokinase*
SUMO protease 12588018 Recombinant fragment of ULP1 from S. cerevisiae that contains 6xHis for easy removal Tertiary structure of SUMO peptide Removal of SUMO fusion tags
Sequence recognized Used for
Asp-Asp-Asp-Asp-Lys Cleavage of any fusion protein containing the recognition sequence
*Use EK-Away Resin (Cat. No. R18001) to remove EKMax Enterokinase (or other enterokinases) from your fusion protein following digestion.
For more information on these products or to place an order, visit lifetechnologies.com/proteases.
Analytical and production chromatography
Weve developed chromatography resins and systems that accommodate protein production at every scale from pilot projects up to process-scale separations. Choose one of our high-resolution, high-capacity analytical and process resins or one of our specialty resins, designed for specific applications such as large and small protein biomolecules, viruses, and antibodies.
Product
POROS Perfusion Chromatography Resin
Application
High performance, high throughput for process-scale bioseparations Separation of biomolecules for both analytical and lab-scale preparation Biopharmaceutical manufacturers in earlyphase process development Microbial protein purification Purification of viruses or virus-like particles for vaccine production Pilot- or production-scale antibody purification
Particle size
50 m 10 and 20 m 20 and 50 m 50 m 50 m 10, 20, or 50 m
Online resource
lifetechnologies.com/porosproduction lifetechnologies.com/porosanalytical lifetechnologies.com/gopure lifetechnologies.com/microbialresins lifetechnologies.com/virusresins lifetechnologies.com/antibodyresins
POROS Analytical Chromatography GoPure Pre-packed Chromatography Columns Resins for microbial expression systems Resins for viral vaccine production Resins for monoclonal antibody production
For more information, visit lifetechnologies.com/proteinchromatography.
131
Protein analysis
Protein sample fractionation
The ZOOM IEF Fractionator reduces highly complex protein samples into fractions, based upon isoelectric point, for analysis by two-dimensional gel electrophoresis (2DE), one-dimensional gel electrophoresis (1DE), or two-dimensional liquid chromatography/mass spectrometry (2D LC/MS). The ZOOM IEF Fractionator offers a range of rapid separation (fractionation) options and:
Allows loading of high amounts of protein for downstream applications Enriches low-abundance proteins and increase the dynamic range of detection Reduces precipitation/aggregation artifacts associated with highprotein load
samples To learn more about the Zoom Fractionator and reagents, visit lifetechnologies.com/ proteinfractionation.
ZOOM IEF Fractionator
To reduce sample complexity prior to analysis, isolate organelles with magnetic separation technology. Dynabeads can been used to isolate a number of different organelles and organelle-specific proteins. The magnetic separation technology enables specific isolations of true fractions in small volumes, and can replace timeconsuming and labor intensive density gradient centrifugations.
Organelle isolation kits
Rapid, high-purity isolations from crude fractions or fractions of common Morphologically intact target organelles can be fixed for EM studies or solubilized Immunomagnetically isolated organelles can be assayed by all conventional Easy protocol yields high-purity functional organelles of interest
Examples of organelles or organelle fractions that have been isolated with Dynabeads products include mitochondria, peroxisomes, kidney glomeruli, and endocytic and secretory compartments. techniques directly on the bead surface densities
Need Isolation of fragile organelles or organelle fractions
Application requirement Mouse primary antibody
Recommended product Dynabeads M-280 Sheep AntiMouse IgG (Cat. No. 112-01D) Dynabeads M-280 Sheep AntiRabbit IgG (Cat. Nos. 112-03D, 112-04D) Dynabeads Protein G (Cat. No.100-03D) Dynabeads Protein A (Cat. No. 100-01D) Dynabeads M-450 with a secondary antibody against the corresponding primary antibody species (Cat. No. 140-13) Dynabeads M-280 Tosylactivated (Cat. No. 142-03) Dynabeads M-270 Epoxy (Cat. No. 143-01)
Advantage Direct or indirect isolation of organelles made more efficient with two layers of antibodies Direct or indirect isolation of organelles made more efficient with two layers of antibodies Efficient direct or indirect isolation of organelles More efficient pull to the magnet in viscous samples Easy coupling of a molecule onto these hydrophobic beads for efficient organelle isolation Coupling of a molecule onto these hydrophillic beads for very high-purity organelle isolation
Rabbit primary antibody
Non-mouse/non-rabbit primary antibody Isolation of large organelle fractions or isolation from a viscous sample Isolation of organelles using a non-antibody ligand Antibodies made to specific species High yield
High purity
To learn more about organelle isolation with Dynabeads or to find products, visit lifetechnologies.com/organelleisolation.
132
Protein analysis
Cell lysis buffers and protein extraction kits
Easy-to-use buffers and kits allow rapid and simple protein extraction as well as protein extraction combined with RNA isolation from cultured cells. The table below lists some of our top lysis and extraction products.
Buffer Cell Extraction Buffer* NP-40 Cell Lysis Buffer* Tissue Extraction Reagent 1* PARIS Kit**
Best for Sample prep of cells prior to ELISA and western blot Sample prep of cells prior to Luminex assays, ELISA, western blot; contains NP-40 detergent Total protein extraction from tissue samples Isolating total RNA and total protein from fresh cultured cells; alternatively, cells can be partitioned into nuclear and cytoplasmic fractions before protein/RNA fractionation; phenol-free formulation; useful for enzymatic assays, immunoprecipitation, gel shift assays, 2D electrophoresis, and western blotting Correlating mRNA, miRNA, or siRNA levels with protein levels; achieves simultaneous extraction of native protein and RNA from cells or tissues
Cat. No. FNN0011 (standard buffer, 100 mL), FNN0091 (denaturing buffer, 100 mL) FNN0021 (100 mL) FNN0071 (100 mL) AM1921 (50 purifications)
mirVana PARIS Kit**
AM1556 (2040 isolations)
* Go to lifetechnologies.com/proteinlysisbuffer for more information. ** Go to lifetechnologies.com/proteinextractionkits for more information.
133
Protein analysis
Qubit 2.0 Fluorometer

Simple, accurate protein quantitation
The Qubit 2.0 Fluorometer together with the Qubit Protein Assay offer an easyto-use method for rapid quantitation of protein samples. The Qubit 2.0 Fluorometer reads the signal from an optimized Molecular Probes dye that is selective for protein even in the presence of an equal mass of DNA or RNA. Qubit fluorometric quantitation is:
Selectivea protein-selective dye in the Qubit Protein Assay has a linear

detection range of 0.255 g of protein (effective for starting sample concentrations from 12.5 g/mL to 5 mg/mL) Sensitivesamples with concentrations as low as 12.5 g/mL of protein may be accurately and reliably quantitated Simple and intuitivethe Qubit 2.0 Fluorometer offers rapid results and is easy to use Features of the Qubit 2.0 Fluorometer include:
The Qubit Fluorometric Quantitation System.
Protein quantitation
Large LCD color touch screen Automatic data logging and USB port for data management Easy workflow navigation Standard curve display after calibration is completion
The Qubit assays for use with the Qubit 2.0 Fluorometer are all performed using the same simple mix-and-read protocol. The Qubit Protein Assay Kits (Cat. Nos. Q33211 and Q33212) provide concentrated assay reagent, 1X buffer, and prediluted BSA standards. Simply dilute the reagent using the buffer provided, add your sample (any volume between 1 L and 20 L is acceptable), incubate for 15 minutes, and read the concentration using the Qubit 2.0 Fluorometer.
A simple protocol gives you quantitation data in minutes
Standards from kit
10 L
10 L
Add Quant-iT working solution for a total volume of 200 L
Vortex tubes for 23 sec Incubate at room temperature for 2 min (DNA, RNA) or 15 min (protein)
User samples
120 L
120 L
120 L
Add Quant-iT working solution for a total volume of 200 L
Read tubes in Qubit Fluorometer
Workflow for the Qubit assay using the Qubit 2.0 Fluorometer.
134
Protein analysis
The Qubit Protein Assay exhibits very low protein-to-protein variability
Compared to other protein assays, the Qubit Protein Assay exhibits very low proteinto-protein variability (see figure). For the most accurate results when quantitating protein with the Qubit Protein Assay, use detergent-free protein samples. To see which contaminants are tolerated by the Qubit Protein Assay, see Table 2 of the Qubit Protein Assay Kits manual. You can download the manual at lifetechnologies.com/qubitproteinassay.
Qubit Protein Assay
Pierce BCA Protein Assay
Protein concentration determined by assay (mg/mL)
0.700 0.600 0.400 0.400 0.300 0.200 0.100 0.000 -0.100 0
BSA Lysozyme Histone Bovine pituitary extract
0.800
0.800 0.700 0.600 0.400 0.400 0.300 0.200 0.100 0.000 -0.100
Protein quantitation
0.1
0.2
0.3
0.4
0.5
0.6
0.1
0.2
0.3
0.4
0.5
0.6
[Protein] (mg/mL)
Bio-Rad Quick Start Bradford Protein Assay
[Protein] (mg/mL)
Pierce Lowry Protein Assay
0.800 0.700 0.600 0.400 0.400 0.300 0.200 0.100 0.000 -0.100 0 0.1 0.2 0.3 0.4 0.5 0.6
0.800 0.700 0.600 0.400 0.400 0.300 0.200 0.100 0.000 -0.100 0 0.1 0.2 0.3 0.4 0.5 0.6
[Protein] (mg/mL)
[Protein] (mg/mL)
Protein-to protein variability. Triplicate samples of BSA, lysozyme, histone, and bovine pituitary extract were assayed at concentrations of 00.5 mg/mL using the Qubit Protein Assay on the Qubit 2.0 Fluorometer, and various other protein quantitation methods. The variability at 0.3 mg/mL for the Qubit assays was found to be <7%.
135
Protein analysis
Gel electrophoresis
High-performance, high-quality precast gels and gel casting essentials

If you are doing standard one-dimensional protein separation, we have Novex solutions, and this section will help you select the separation tools that fit your research. The Novex gel line consists of several different chemistries, well formats, and gel sizes, so you get the separation you need for accurate downstream results. To learn more about our basic denaturing protein separation, visit lifetechnologies.com/buyproteingels.
One-dimensional protein separation.

Gel-casting accessories Application Pour-your-own essentials: preassembled empty cassettes, buffers, reagents Medium Medium NA 90 min 6500 kDa NA Tris-glycine gels Precast convenience with Laemmli chemistry NuPAGE gels Straight protein bands, longest shelf life Bolt gels Optimized for western blot performance
Protein gel electrophoresis
Protein resolution Protein integrity Shelf life Average run time Separation range Gel % available
Medium Medium 12 months 90 min 6500 kDa 4%, 6%, 8%, 10%, 12%, 420%, 412%, 14%, 16%, 18%, 816%, 1020% 5- and 10-packs Mini: 8 x 8 cm, 1 or 1.5mm thick Midi: 8 x 13 cm, 1mm thick 25 L (1 mm) 37 L (1.5 mm) 0.5 g
High High 8 months (Tris-acetate) 16 months (Bis-Tris) 35 min 1.5300 kDa (Bis-Tris) 30400 kDa (Tris-acetate) 412%, 8%, 10%, 12% (Bis-Tris) 38%, 7% (Tris-acetate) 2- and 10-packs Mini: 8 x 8 cm, 1 or 1.5mm thick Midi: 8 x 13 cm, 1mm thick 30 L (1 mm) 43 L (1.5 mm) 0.5 g
High High 16 months 35 min 1.5300 kDa 412%, 8%, 10%, 12%
Pack sizes available Gel dimensions
NA Mini: 8 x 8 cm, 1 or 1.5mm thick Midi: 8 x 13 cm, 1mm thick 25 L (1 mm) 37 L (1.5 mm) NA
10-pack 8 x 8 cm, 1 mm thick, with wedge-shaped wells
Maximum sample volume/well Maximum protein load per band for optimal resolution*
60 L 0.5 g
*On 1.0 mm, 10-well gel, detected by Coomassie staining.
136
Protein analysis
Bolt gels
Bolt Bis-Tris Plus Gels are optimized to deliver superior western blot performance:
Superior band quality and band volumeattributed to Novex Bis-Tris Plus Better protein resolution with more protein bands detecteddue to 10% Preserved protein integrity with the legendary Novex neutral-pH formulation Best lot-to-lot consistencyour best-in-class manufacturing delivers highly
reproducible gel performance, with CV of only 2% for Rf resolution values Bolt gels are designed to deliver optimal resolution, band quality, and sensitivity for western blot analyses. Bolt gels give well-resolved, flat, straight western blot bands compared to other commercially available gels that present rounded, poorly resolved bands. Besides providing more accurate molecular weight assignments of your proteins, Bolt gels overall provide better-looking, publication-quality western blots. Unlike traditional Tris-glycine gels, Bolt Bis-Tris Plus gels are Bis-Tris HCl buffered (pH 6.4) and have an operating pH of about 7.0. This neutral pH paired with a unique, gentle sample preparation protocol means your western protein samples always experience mild, nonacidic conditions. This preserves protein integrity and minimizes protein modifications to help ensure highly sensitive, accurate western blots every time. For more information and to place your order, visit lifetechnologies.com/bolt. that minimizes protein modifications
Bolt Bis-Tris Plus Gels.
chemistry for delivery of sharp, straight bands
greater resolving distance and optimized gradient format
Western blot band quality from Bolt gels. A Bolt 412% gradient gel was run at 165 V for 35 min. Two GST fusion proteins as well as GST were spiked into E. coli lysate (0.6 mg per gel lane) to create series of decreasing, increasing, or constant amounts of these proteins across the gel. The spiked amounts of the two fusion proteins ranged from 40 ng to 0.2 ng per gel lane. A WesternBreeze Chemiluminescence Detection Kit was used with an anti-GST antibody. The image was acquired using an LAS-1000 (FujiFilm) system.
137
Protein analysis
Wedge well for loading up to 2x more protein sample
Bolt gels have new wedge-shaped wells for loading up to twice as much protein sample in every well. The new wedge well: Allows loading of dilute samples Minimizes sample spillover and cross-well contamination Enables easy gel loading with standard pipette tips Delivers better detection sensitivity for low-abundance proteins Gives better stacking of high molecular weight proteins to give better resolution Enables greater success with low-affinity western antibodies For more information and to place your order, visit lifetechnologies.com/bolt.
Wedge wells on the Bolt Bis-Tris Plus Gel.
NuPAGE gels Protein gel electrophoresis
The NuPAGE precast gel system is a revolutionary high-performance polyacrylamide gel system.
Superior protein band resolution and stability Faster sample run times Longer product shelf life
Gels are available in both Bis-Tris and Tris-acetate formulations and in a variety of acrylamide percentages (see table). The unique NuPAGE gel formulations minimize the smiles and poor resolution seen with other gel types. The neutral-pH gel chemistry and buffer system avoid chemical modifications to the samples that occur in alkaline environments. In addition, proteins transfer from NuPAGE gels more efficiently, resulting in higher detection signals during western analysis. For more information on our NuPAGE products and to place an order, visit lifetechnologies.com/nupage.
Novex NuPAGE 412% Bis-Tris Midi Gel.
Two NuPAGE types to choose from.

Gel type Bis-Tris Percentages available 8%, 10%, 12%, 412% Separation range 1.5300 kDa Shelf life 16 months* Average run time 3550 min Applications Separation of small- to medium-sized proteins; simple sample reduction; protein sequencing, mass spectrometry, techniques where protein integrity is crucial Excellent separation of large molecular weight proteins
Tris-acetate
7%, 38%
30400 kDa
8 months*
60 min
138
Protein analysis
Convenient electrophoresis
Intuitive electrophoresis tank The Bolt Mini Gel Tank (Cat. No. B4477599) is designed for more intuitive use and convenience than traditional electrophoresis tanks:
Easy sample loadingwith the new front well configuration Less running buffer requiredwith two separate gel chambers, you only need to load sufficient buffer Simultaneous visualization of both gelswith streamlined, side-by-side tank configuration Simplified monitoring of prestained markerswith new white tank stand
For more information and to place your order, visit lifetechnologies.com/bolt. for each gel
Bolt Mini Gel Tank.
All Novex gels are available in the 8 x 8 cm mini gel format and a larger 8 x 13 cm midi size for higher throughput. Run up to two mini gels at a time in the XCell SureLock Mini-Cell (A, below). Use the XCell4 SureLock Midi-Cell (B, below) to simultaneously run up to four midi gels. Each midi gel can run up to 26 samples, allowing electrophoresis of over 100 samples at a time. With a Midi Gel Adapter (Cat. No. WA0999), Novex Midi Gels can be run in a Bio-Rad Criterion Cell. For more info on these and other products for running Novex gels and to place an order, visit lifetechnologies.com/electrophoresisaccessories.
Gel tanks and buffers
The XCell SureLock protein electrophoresis cells. (A) XCell SureLock Mini-Cell (Cat. No. EI0001) for running one or two NuPAGE or Novex mini gels. (B) XCell4 SureLock Midi-Cell (Cat. No. WR0100) for running up to four NuPAGE or Novex midi gels.
139
Protein analysis
Additional gels from Novex brands
The Novex family of products offers a variety of options for protein separation, including gels for isoelectric focusing, resolution of undenatured proteins, low molecular weight proteins and even 2 dimension electrophoresis. Please review the selection table below to find the gel most suited for your research needs. For more information on our comprehensive line of specialty gels for protein separation and to place an order, visit lifetechnologies.com/specialtygels.
Specialty protein gels

Quickly analyze 4896 proteins E-PAGE gels Analyze posttranslational protein modifications Novex IEF gels Excellent resolution for low MW Novex Tricine gels Separate low-MW proteins and peptides 220 kDa 25 L Optimized resolution of active proteases Novex zymogram gels Separate proteases based on size and activity 10220 kDa 20 L Superior resolution of native proteins NativePAGE gels Convenient and simplified 2D electrophoresis ZOOM 2D Electrophoresis System Fractionate proteins before electrophoresis and MS pH 312 140 L (diluted)
Product
Applications
Compatible with high-throughput systems 10200 kDa 20 L (E-PAGE 48) 15 L (E-PAGE 96)
Separate native proteins and study modifications pI 3.56.8 pI 3.58.5 20 L
Determine size and purity of native proteins 1510,000 kDa 25 L
Separation Range Max sample volume/well
Gel % available
8% (E-PAGE 48)
5%
10%, 16%, 1020%
10% (with gelatin)
312%, 416%
pH 310 NL, 310L, 47, 610, 912 (broad range) pH 4.55.5, 5.36.3, 6.17.1 (narrow range)
6% (E-PAGE 96)
12% and 416% (with casein) 2.5 hrs 90 min 90 min 90 min
Average run time
14 min
30 min (1st dimension) 40 min (2nd dimension)
Shelf life Gel dimensions
6 months 13.5 x 10.8 cm, 3.7mm thick
2 months 8 x 8 cm, 1.0mm thick
12 months 7 cm strips, 0.5cm thick
140
Protein analysis
Accurate molecular weight estimations

Sharp, accurate, prestained and unstained standards
Protein separation is often accompanied by band identification. A variety of Novex molecular weight (MW) protein standards are available to help you make the most accurate size estimations possible.
Sharpest bandsclearer estimation of MW compared to thick bands Broadest MW rangeestimation over a larger range (3.5260 kDa) Ladder-like separation of bandsevenly spaced pattern across the readable range
for easy identification and comparison To learn more about protein molecular weight standards and to place an order, go to lifetechnologies.com/proteinstandards.
Novex prestained standards. Find this interactive table online at lifetechnologies.com/prestainedproteinstandards.

SeeBlue Pre-stained Standard (Cat. No. LC5625) Application Basic, routine MW BenchMark HiMark Pre-stained Pre-stained Protein Protein Standard Ladder (Cat. No. 10748010) (Cat. No. LC5699) MW marker for Tris-glycine gels only 10 bands 2 colors Low 10190 kDa Tris-glycine only
SeeBlue Plus2 Pre-stained Standard (Cat. No. LC5925) Multiple color bands, easy estimation 10 bands 3 colors Medium 4250 kDa NuPAGE Bis-Tris; Tricine, Tris-acetate, Tris-glycine
Sharp Pre-stained Protein Standard (Cat. No. LC5800) Easiest to interpret MW; best size estimation 12 bands 12 colors High 3.5260 kDa NuPAGE Bis-Tris; Tricine, Tris-glycine
Ideal for large proteins 9 bands 2 colors Low 30460 kDa Tris-acetate, Trisglycine
Number of bands and colors Band sharpness MW range Gel compatibility
9 bands 1 color Low 4250 kDa NuPAGE Bis-Tris; Tricine, Tris-glycine
Unstained protein standards. Find this interactive table online at lifetechnologies.com/unstainedproteinstandards.

Mark12 Unstained Standard (Cat. No. LC5677) Application Routine MW estimation in denaturing gel electrophoresis 12 2.5200 kDa BenchMark Protein Ladder (Cat. No. 10747012) MW estimation and quick orientation Sharp Unstained Protein Standard (Cat. No. LC5801) MW estimation and band identification HiMark Unstained Protein Standard (Cat. No. LC5688) MW estimation for large proteins NativeMark Unstained Protein Standard (Cat. No. LC0725) MW estimation in native gel electrophoresis
Number of bands MW range
15 10220 kDa Yes 2 (20 and 50 kDa bands more prominent) NuPAGE Bis-Tris; SDS-PAGE Tricine, Tris-acetate, Trisglycine Medium
12 3.5260 kDa Yes 3 (20, 50, and 110 kDa bands more prominent) NuPAGE Bis-Tris; SDS-PAGE Tricine, Tris-acetate, Trisglycine High
9 40500 kDa No 0
8 201,200 kDa No 0
Orientation band(s) No Number of 0 orientation band(s) Gel compatibility NuPAGE Bis-Tris; SDS-PAGE Tricine, Tris-acetate, Trisglycine Low
Tris-acetate
Tris-glycine, NativePAGE Bis-Tris, Tris-acetate Low
Band sharpness
Low
141
Protein analysis
Sensitive stains for total detection

Life Technologies offers a number of staining reagents for total protein and posttranslationally modified proteins, including Coomassie G-250 stain, silver stain, and a suite of compatible and highly sensitive fluorescent stains that allow direct quantitation of differential phosphorylation and glycosylation patterns as well as total-protein expression from a single sample on the same gel. Use the table below to find the stain thats right for your application.
Overview of Novex protein stains. For more information on protein gel stains, visit lifetechnologies.com/proteinstains.
Product Cat. No. Type of stain Instrumentation required SimplyBlue SafeStain LC6060, LC6065 SilverQuest Silver Staining Kit LC6070 SYPRO Ruby Protein Gel Stain S12001, S12000, S21900 Fluorescent; ready-to-use UV transilluminator, bluelight box, or laser scanner Subnanogram Low 0.251,000 ng Yes Integration of fluorescent signal Yes Use SYPRO Ruby Protein Blot Stain 90 min (mini gels with new protocol); overnight for large-format gels Fix, stain, and wash Coomassie Fluor Orange Protein Gel Stain C33250, Fluorescent; ready-to-use UV transilluminator, bluelight box, or laser scanner 48 ng Low 41,000 ng No Integration of fluorescent signal Yes No 3060 min
Colorimetric; ready-to-use Colorimetric; colloidal Coomassie G-250 ready-to-use silver No >7 ng BSA Low 770 ng No Yes Yes PVDF 3 hr; 12 min with microwave protocol Rinse, stain, water-based destain No Subnanogram Low 110 ng No Not recommended Yes No 90 min; <60 min with microwave protocol 10 solution changes
Sensitivity Background Linear dynamic range Multiplex capability For densitometry MS compatibility Staining membranes Total staining/ destaining time Protocol summary
Fix and stain in one step
SYPRO Ruby protein gel stain for ultrasensitive total protein detection
Simple staining procedure Linear quantitation range of 3 orders of magnitude Linear compatibility with mass spectrometry and microsequencing
SYPRO Ruby Protein Gel Stain is a highly sensitive fluorescent stain for proteins in 1D or 2D gels (see figure). The fast microwave mini gel protocol yields optimal results, typically in only 90 minutes. For convenience, SYPRO Ruby stain is provided ready to use and stores at room temperature. Learn more at lifetechnologies.com/proteinstains.
Sensitive fluorescent staining with SYPRO Ruby Protein Gel Stain.
142
Protein analysis
Efficient protein transfer for successful western blotting

iBlot 7-Minute Blotting System
Complete transfer in 7 minutes or less High blotting efficiency and evenness Increased blotting reliability and reproducibility
The iBlot 7-Minute Blotting System (Cat. No. IB1001) is designed to efficiently and reliably blots proteins from polyacrylamide gels in 7 minutes or less, without the need for buffers or an external power supply. A self-contained unit, the iBlot device uses disposable blotting stacks with integrated nitrocellulose or PVDF transfer membranes, offering the convenience of a bufferless, plug-and-play system. In addition to ease and convenience, the iBlot system offers highefficiency protein transfers (see figure) for high downstream detection sensitivitiesin many cases, higher than conventional methods. In the end, you can achieve more accurate detection using less sample. The iBlot 7-Minute Blotting System is designed to work with multiple polyacrylamide gel sizes, including the midi size, fitting midi gels (8 x 13 cm) and E-PAGE 48 and 96 gels, and mini gels (8 x 8 cm). A
1 2 3 4 5 6 7 8 9 10 11 12
The iBlot 7-Minute Blotting System.
B
1 2 3 4 5 6 7 8 9 10 11 12
High transfer efficiencies achieved using the iBlot 7-Minute Blotting System. (A) iBlot dry transfer to nitrocellulose and (B) semi-dry transfer to nitrocellulose of NuPAGE 12% Bis-Tris Mini Gels. Lanes 16: 0.0625 g, 0.125 g, 0.25 g, 0.5 g, 1.0 g, and 2.0 g of SW480 human colon cancer cell lysate (Alexis) (control); lanes 7 and 12: 5 L SeeBlue Plus2 Pre-stained Protein Standard; lanes 811: 0.5 L, 1.0 L, 2.0 L, and 4.0 L of MagicMark XP Western Protein Standard.
Which western transfer system is right for you?
Application Product Transfer time Capacity of device Blotting area Requires transfer buffer Power supply
Traditional wet transfer in 12 hr XCell II Blot Module (Cat. No. EI9051) 60120 min 12 mini gels 9 x 9 cm Yes (1,000 mL) External
Semi-dry transfer for high efficiency Novex Semi-Dry Blotter (Cat. No. SD1000)
Dry transfer in 7 min iBlot Gel Transfer Device (Cat. No. IB1001) 7 min 2 mini gels or 1 midi gel 13.5 x 8.5 cm No Internal
3060 min 4 mini gels or 2 midi gels 21 x 21 cm Yes (500 mL) External
For more information and to place an order for western transfer systems, visit lifetechnologies.com/westerntransfer.
143
Protein analysis
Precut blotting membranes
Simplify blotting setup with precut membrane/filter-paper sandwiches. Life Technologies makes blotting easier by providing a variety of precut, preassembled membrane/ filter-paper sandwiches for mini and midi/E-PAGE gels (see figure). A proteins properties (i.e., charge, hydrophobicity, etc.) affect its ability to bind to membrane surfaces. Finding the right membrane may require experimenting with your specific protein on different membranes. For more information about our western analysis products, go to lifetechnologies.com/iblot.
1 2 3 4 1 2 3 4
Western transfer and detection
Invitrolon PVDF
Other 0.45 m PVDF
High signal achieved on Invitrolon PVDF. A 53 kDa protein containing a c-Myc epitope was transferred to (A) Invitrolon PVDF and (B) another manufacturers 0.45 m PVDF membrane. Both PVDF membranes were probed with a 1:500 dilution of mouse anti-Myc antibody, then developed with the WesternBreeze Chemiluminescent Anti-Mouse Kit. Blots shown here are 2 min exposures on X-ray film. Lanes 14: 2 ng, 1 ng, 0.5ng, and 0.2 ng of protein.
BenchPro 4100 Western Processing System

Just push run and walk away
Reduces tedious hands-on work and human errors Minimizes cross-contamination, and no clean-up Automate without protocol changes Compatible with all chemiluminescent, chromogenic, and fluorescent immunodetection reagents and protocols. The BenchPro 4100 Western Processing System (Cat. No. WP0001) helps consistently and accurately deliver the correct volumes of solutions to the membrane at precise times. This automation removes the tedious and repetitive liquid handling steps associated with manual western blot processing. Western blots processed using the BenchPro 4100 system minimize human processing errors, so you get greater experimental reproducibility and data consistency.
BenchPro 4100 Western Processing System.
4 B
The BenchPro 4100 Western Processing System is designed to generate consistent, reproducible results, run after run. It is compatible with all chemiluminescent, chromogenic, and fluorescent immunodetection reagents and protocols (see figure). The BenchPro 4100 system dramatically reduces the need for protocol optimization and allows you to perform immunoblotting your way, right away. For more information, go to lifetechnologies.com/benchpro4100.
Manual processing vs. BenchPro 4100 processing of western blots. (A) Manual processing. (B) BenchPro 4100 system processing. Lane1: 8 L of a 1:10 dilution of MagicMark XP Standard; lane 2: 50 ng BSA; lane 3: 25 ng BSA; lane 4: 10 ng BSA. Proteins were detected using rabbit anti-BSA antibody and WesternBreeze Chemiluminescent KitAnti-Rabbit. Detection substrate was added to both membranes, and the blots were imaged at the same time.
144
Protein analysis
Novex western blot detection kits

Life Technologies offers a broad range of reagents and kits for western blot detection. Whether you are using chromogenic, fluorescent, or chemiluminescence detection systems, we have the solutions you need.
iBlot Western Detection Kits
Fastcomplete western detection, typically in less than 25 minutes when used

with the iBlot 7-Minute Blotting System Flexibleworks with mini and midi gels Easy optimizationallows different conditions to be applied to different sections of the blot The iBlot Western Detection Kits (see figure) consist of iBlot detection stacks and iBlot detection reagents. The kits are available with anti-mouse or anti-rabbit secondary antibodies and are compatible with chemiluminescent and chromogenic detection. The iBlot Western Detection Kits offer comparable or better sensitivity than the conventional protocols for the majority of the antibody-antigen pairs, while offering significant time savings. For more information and to place an order, visit lifetechnologies.com/westerndetection.
Comparison of an iBlot Western Detection Kit to a WesternBreeze kit. Proteins from an SW480 (human colon adenocarcinoma cell line) lysate were transferred using the iBlot 7-Minute Blotting System. A mouse anti-p53 antibody was used as the primary antibody. Detection was performed with (A) the WesternBreeze Chemiluminescent KitAnti-Mouse or (B) the iBlot Western Detection, Chemiluminescent Kit (AntiMouse). The iBlot western detection kit detected the proteins with sensitivity comparable to the WesternBreeze kit.
Western transfer and detection
Which western blot detection kit is right for you?
Application Product recommended
Sensitive chemiluminescent detection Novex chemiluminescent substrates ~3 hr Picogram Chemiluminescent HRP No No Film developer/scanner
Fluorescence-based kits utilizing Qdot technology WesternDot 625 Western Blot Kits ~3 hr <1 ng Qdot fluorescent technology Biotin Yes Yes Blue or UV transilluminator
Sensitive, completekits WesternBreeze Western Blot Immunodetection Kits ~3 hr Femtogram to picogram Chemiluminescent or chromogenic AP Yes Yes Film developer/scanner
Sensitive western detection in 30minutes iBlot Western Detection Kits < 30 min Femtogram to picogram Chemiluminescent or chromogenic AP Yes Yes iBlot Gel Transfer Device
Protocol time (not including transfer) Sensitivity Detection method Compatible secondary antibody conjugates Blocking and wash solutions included Secondary antibody included Instruments required
145
Protein analysis
Novex primary antibodies for western blotting

Designed for specific, high-avidity binding to the target of interest Superior lot-to-lot consistency and excellent signal-to-noise ratios Exhibit excellent performance in a variety of immunodetection applications
Life Technologies offers high-quality, highly specific primary antibodies for a broad range of applications and targets, combining the breadth and quality of Invitrogen, Zymed, Caltag, and Biosource under one brand: Novex. Our primary antibodies are created in-house or licensed from leading scientists from around the world, purified using methods designed to retain affinity and ensure stability, and tested in multiple applications such as western blotting, immunohistochemistry, immunofluorescence, flow cytometry, ELISA, neutralizing assays, and other functional studies.
Primary antibodies for western
Phospho- and cleavage-sitespecific antibodies Total (pan) antibodies Cytokine, chemokine, and growth factor antibodies -amyloid, tau, and other neuroscience antibodies CD antibodies Cell junction (gap and tight) antibodies Mitochondrial oxidative phosphorylation system antibodies Epigenetic antibodies Epitope tag antibodies Stem cell antibodies
Our selection of antibodies covers a diverse range of research areas, including (but not limited to):
Quickly and easily find exactly the right primary antibody
Life Technologies offers over 2,800 antibodies, and we are adding new ones every day. For the fastest and easiest way to find the primary antibody you need, use the Primary Antibody Search Tool (lifetechnologies .com/antibodies). Simply search by target, gene symbol, or gene ID, then filter your results by application, reactivity, host, conjugate type, or antibody type. You can also search within your results to quickly find the right antibody.
146
Protein analysis
ABfinity recombinant monoclonal antibodies
Engineered antibodies for consistent results
High performance with proven lot-to-lot consistency Undergo rigorous validation to help ensure specificity Enables you to detect low-level targets with less sample
Produced using our proprietary technology, ABfinity recombinant rabbit monoclonal antibodies are highly specific, high-quality antibodies, superior in consistency and performance. ABfinity recombinant antibodies are produced on a large scale by expressing them in mammalian cells, and appear just like their counterparts isolated from serum or produced by hybridomas. Intact IgG appears on a nonreducing gel as ~150 kDa band and upon reduction generates a ~25 kDa light chain and a ~50 kDa heavy chain. Find out about the quality and consistency of ABfinity antibodies at lifetechnologies.com/abfinity. Lot-to-lot consistency Each ABfinity antibody is manufactured in mammalian cells with antibody heavy and light chain cDNAs from a functionally screened, immunogen-specific antibody. This highly reproducible process results in superior lot-to-lot consistency, which helps save time and money by minimizing or eliminating the need to revalidate performance on new lots. The figure (right) shows the consistent western blotting results achieved using independent lots of an ABfinity antibody. Reliable sensitivity and specificity The ABfinity technology delivers highly sensitive and specific antibodies that only react with the target of choice, minimizing detection of the wrong signal due to nonspecific binding. In addition, high-affinity antibodies allow you to use less antibody for detection, detect very low-level targets, and conserve your precious samples. This western blot (right) shows a direct comparison of an ABfinity STAT4 antibody with the best commercial STAT4 antibodies in western blotting. This comparison includes antibodies from polyclonal, traditional hybridoma monoclonal, and rabbit hybridoma monoclonal platforms. Extensive validation and characterization ABfinity antibodies are validated and characterized by multiple applications. The figures on the next page are the results of an individual antibody in multiple applications: flow cytometry, immunocytochemistry, immunohistochemistry, and western blot. This extensive validation process allows confidence in target specificity, without any need for optimization.
SMAD2
Lot 1 kDa 250 A B 150 100 75
Lot 2 C A B C A
Lot 3 B C A
Lot 4 B C
50 37 25 20 15
Lot-to-lot consistency. HeLa cell extracts were separated on reducing gels, blotted, and probed with four lots of SMAD2 ABfinity Antibody. Lanes A, B, and C were loaded with serial dilutions of the HeLa extract. The primary antibody was detected using the WesternBreeze Chemiluminescent Kit Anti-Rabbit.
kDa
150 100 75 1
AB nity antibody 70185 2 3
Polyclonal antibody 714500 4 5 6
Competitor A 7 8 9
Competitor Competitor B C 10 11 12 13 14
50 37 25 20 15
Reliable sensitivity and specificity. STAT4 ABfinity and polyclonal antibodies were used at 2, 1, and 0.5 g/mL. STAT4 antibodies from other vendors were used at their recommended concentrations. The primary antibody was detected using the WesternBreeze Chemiluminescent Kit Anti-Rabbit.
147
Protein analysis
250
1 kDa 150 100
Number of events
200
150
75 60 kDa 50
100 50
37
100 101 102 103 104
25
Fluorescence
Immunohistochemistry of human esophagus carci noma tissue labeled with AKT [pS473] ABfinity Recombinant Rabbit Monoclonal Antibody. Formaldehyde-fixed, paraffin-embedded (FFPE) human esophagus carcinoma tissue was labeled with rabbit antiAKT [pS473] (0.5 g/mL). Tissues were pretreated with EDTA and detected with SuperPicture Polymer DAB. Image was taken at 20x magnification. Note nuclear and cytoplasmic staining in tumor cells.
Flow cytometry of Jurkat cells labeled with AKT [pS473] ABfinity Recombinant Rabbit Monoclonal Antibody. Jurkat cells were incubated with 50 M PI3K/AKT signaling pathway inhibitor LY294002 (red trace) or without (green trace) for 1 hr prior to being fixed and permeabilized using FIX & PERM reagents. Cells were then stained with 1 g/test of ABfinity Recombinant Rabbit Monoclonal Antibody followed by Alexa Fluor 488 goat antirabbit IgG. The blue trace represents secondary antibody alone.
Western blot of 3T3 cell lysates labeled with AKT [pS473] ABfinity Recombinant Rabbit Monoclonal Antibody. Rabbit antiAKT [pS473] (0.1 g/mL) was used to label AKT [pS473] in untreated 3T3 lysates (lane 1) or PDGF-treated 3T3 lysates (lane 2).
Immunocytochemistry of mouse fibroblasts cells labeled with AKT [pS473] ABfinity Recombinant Rabbit Monoclonal Antibody. Mouse fibroblast cells were treated with (A) or without (B) 10 g/mL insulin and labeled with rabbit anti-AKT [pS473] (5 g/mL). Signal is knocked down after incubation with the phosphopeptide used as antibody (C) but not with the nonphosphopeptide (D). Alexa Fluor 488 goat antirabbit IgG at 1:1,000 was used as secondary antibody. Nuclei are stained with Hoechst (blue).
Recombinant proteins
We offer a wide selection of Novex recombinant proteins for immunology, cancer, and stem cell research.
Affordablechoose the size and price that meet your needs and your budget Activepure, and tested for biological performance Relevantvast portfolio of mammalian proteins suited for your research
Find more information online On our website youll find thousands of recombinant proteins, the majority produced in mammalian systems. Mammalian cell expression ensures proper folding, glycosylation, gamma carboxylation and other post-translational modifications that make the protein more physiologically relevant. Proven performance and comprehensive validation data for our proteins ensures that you have confidence in your research. Through a partnership with Sino Biologicals, we are now able to offer you our widest selection of membrane, internal and secreted recombinant proteins, all at extremely competitive prices. Find highly relevant proteins for the study of cancer, immunology, stem cells and more. Go to lifetechnologies.com/recombinantproteins to find: An easily searchable protein product guide Data demonstrating the purity, biological activity, and functionality of our recombinant proteins and growth factors The most popular recombinant proteins and their uses
148
Protein analysis
Protein detection
For sensitive, specific detection of intracellular or extracellular proteins, Life Technologies offers a wide range of Novex immunodetection-based products in convenient, ready-to-use formats. We have developed antibody pair kits and ELISA kits for singleanalyte analysis and Novex multiplex assays for multi-analyte analysis. You can use our immunoassays to investigate: Targets Species Sample types
Cytokines Chemokines Signaling proteins Receptors Neurobiology markers Growth factors Adhesion molecules
Human Mouse Rat Monkey Swine Bovine
Serum Plasma Supernatant Cell lysate Tissue homogenate Urine Cerebrospinal fluid
Go to lifetechnologies.com/immunoassay to find: Information on all of our antibody pair kits, ELISA kits, and Novex multiplex assay kits and instruments Immunoassay Selection Guide that allows you to search for assays based on your target protein Data demonstrating assay specificity and sensitivity
Protein detection
Characteristics of our antibody pair kits, ELISA kits, and Luminex Assay Kits. Go to lifetechnologies.com/immunoassay to find an interactive guide that allows you to search by your protein of interest, then filter by validated application, reactivity, or assay type to find exactly what you need.
Immunoassay type Ready-to-use reagents Analytical sensitivity Dynamic range Incubation time Multiplexable Number of targets measured/well Readout Instrumentation needed Antibody Pair Kit No; need overnight coating process <10 pg/mL 5250 pg/mL 4 hr No 1 HRP-TMB (colorimetric) Microplate reader ELISA Kit Yes <10 pg/mL 5250 pg/mL 2.54 hr No 1 HRP-TMB (colorimetric) Microplate reader Novex Multiplex Assay Kit Yes <10 pg/mL 52,000 pg/mL 3.5 hr Yes 150 R-PE (fluorescent) Luminex instrument (Luminex 100/200, MAGPIX, or FLEXMAP 3D System) 2060 min
Instrument read time
2 min
2 min
149
Protein analysis
Antibody pair kits

Novex Antibody Pair Kits contain matched, pretitered, and fully optimized capture (coating) and detection antibodies. These kits enable you to build your own ELISA or any other assay platform that utilizes a matched antibody pair. For convenience, we also offer a Buffer Kit for Antibody Pairs that contains premade, easy-to-use buffers and solutions that are optimized for use with Antibody Pair Kits.
Convenient format for maximum flexibility Cost savings over ready-to-use ELISA kits Easy to use with optimized reagents and protocol
Antibody Pair Kit supplies sufficient reagents for 10 ELISA plates (5 plates for intracellular targets) and includes: Capture antibody Detector antibody Recombinant standard Streptavidin-HRP conjugate Assay Buffer Set (Cat. No. CNB0011) supplies sufficient reagents for 10 ELISA plates and includes: Coating buffer A Coating buffer B Assay buffer (5X) Wash buffer (25X) Stabilized chromogen (substrate) Stop solution
Protein detection
Learn more at lifetechnologies.com/buildyourownimmunoassay.
ELISA kits
Novex ELISA kits allow specific, quantitative measurements of proteins including cytokines, chemokines, beta amyloids, and signaling targets. Novex ELISA test kits come ready to use with a precoated 96-well plate and all necessary reagents. A detailed, easyto-follow protocol with kit-specific performance information is also provided. Just add sample, run the assay, and typically get quantitative results in half a day.
Specifications of Novex ELISA kits.

Type of ELISA Analytical sensitivity Dynamic range Incubation time Readout Instrumentation needed Colorimetric ELISA <10 pg/mL 5250 pg/mL 4 hr HRP-TMB (colorimetric) Microplate reader Ultrasensitive ELISA <1 pg/mL 0.520 pg/mL 45 hr HRP-TMB (colorimetric) Microplate reader ChemiELISA <1 pg/mL 0.52,000 pg/mL 3.5 hr AP-CSPD (chemiluminescent) Microplate reader with chemiluminescence capability
150
Protein analysis
We research each target protein and calibrate our ELISA kits to provide physiologically relevant sensitivity. In addition, kits are validated using common sample types including serum, plasma, supernatant, and cell lysates for signaling or phosphorylation proteins. Our ELISA kits are manufactured to help ensure excellent quality and reproducibility. Kits must meet quality-controlled specifications for sensitivity, dynamic range, precision, specificity, recovery, and lot-to-lot consistency in order to leave our facilities. Our stringent quality specifications require that each new lot of ELISA kits exhibits minimal variation compared to previous lots.
Rigorous assay validation of Novex ELISA kits helps ensure consistent, reliable results.
Specification Standard calibration Precision Sensitivity Specificity Recovery Lot-to-lot consistency Linearity of dilution Parallelism Description Calibrated to NIBSC, if available Inter-assay CV <10%, intra-assay CV <10% Relevant levels of protein Cross-reactivity tests are performed with similar analytes Buffers are optimized to minimize matrix effects In-house controls are tested to measure within set ranges Linear results over the quantitative range of the assay Recombinant protein standards mimic native proteins What does it mean? Widely used for accurate quantitation and consistent standard of reference Consistent results each time Enables detection of low levels of proteins Helps to ensure accurate measurement of protein of interest Helps to ensure accurate quantitation in serum and plasma Helps to ensure consistent results with new lots Serial dilution of samples quantitate accurately Samples can be measured with confidence
Protein detection
Sensitive, accurate, and consistent performance Validated on serum, plasma, tissue culture supernatant or lysate Ready-to-use, convenient assay in only half a day
Learn more about our standard, ultrasensitive, and chemiluminescent ELISA kits at lifetechnologies.com/elisakits.
Chemiluminescent ELISA kits
High sensitivityallows measurement of proteins expressed at low levels in serum and plasma Large dynamic rangebroad assay range minimizes guesswork of sample dilutions Consistent resultsimproved precision at low end of quantitative range
10,000
Chemiluminescent ELISA Ultrasensitive ELISA
Signal to noise
1,000
Standard ELISA
100
10
Standard curve comparison of Hu IL-1B Chemiluminescent ELISA Kit (Cat. No. KHC0019), Hu IL-1B Ultrasensitive ELISA Kit (Cat. No. KHC0014), and the standard Hu IL-1B ELISA Kit (Cat. No. KHC0011). The chemiluminescent kit has a greater dynamic range than other formats. In addition, the sensitivity levels of the kit is <1 pg/mL, almost 10-fold greater than standard ELISA kits.
0.1
10
100
1,000
10,000
Concentration (pg/mL)
To see specific kits, go to lifetechnologies.com/chemielisas.
151
Protein analysis
Neuro ELISA kits
ELISA kits for accurate A (-amyloid), tau, and -synuclein protein quantitation to assist Alzheimers disease researchers
2.4 2.0 1.6
Easy-to-run sandwich ELISA Precoated, breakable 96-well plate Consistent, accurate, and sensitive measurements
Learn more about our neuro ELISA kits at lifetechnologies.com/neuroelisas.
OD
1.2 0.8 0.4 0 0 600 1,200 1,800
Serial dilution of human CSF containing A1-42 demonstrates linearity over the range of the assay (Cat. No. KHB3441). Linear regression analysis of samples versus the expected concentration yielded a correlation coefficient of 0.99.
Cerebrospinal fluid Dilution 1:4 1:8 1:16 1:32 Measured (pg/mL) 341.0 185.4 94.6 41.2 Expected (pg/mL) 341.0 170.5 85.3 42.6 Expected % 108 107 97
Typical 7-point standard curve for Tau ELISA Kit (Cat. No. KHB0041). The dynamic range is 312,000 pg/mL, and the analytical sensitivity <12 pg/mL.
Protein detection
phosphoELISA kits
4.0
Specificitytwo antibodies directed against analyte provides better specificity than

western blotting Sensitivitymore sensitive than western blotting Sample volumerequires less sample compared to western blotting Quantitationget quantitative data (unlike with western blots) Medium throughput96-well format, results in 4 hours, no densitometry analysis needed Learn more about our phosphoELISA kits at lifetechnologies.com/phosphoelisas. B
OD 450 nm
3.0
2.0
1.0
0.63 1.25
2.5
10
20
40
Protein (g)
Quantitative data from (A) STAT5a [pY694] phospho ELISA assay (Cat. No. KHO0761) confirms that from (B) western blotting using a NuPAGE Novex gel. Assays were performed in parallel.
152
Protein analysis
Move up to multiplexing
Multiplexing allows simultaneous analysis of multiple markers in one sample to measure cytokine secretion and the associated downstream phosphorylated proteins to help give you the complete story both inside and outside the cell. Life Technologies offers a complete solution for the Luminex xMAP technology including polystyrene and magnetic assay kits, reagents, xPONENT systems software, technical support and the entire Luminex portfolio of instruments including the Luminex 200, FLEXMAP 3D, and MAGPIX platforms, covering the full range of multiplexing capabilities. Find more information online We have a wealth of information for you online at lifetechnologies.com/luminex. There youll find: Information about how Novex multiplex assays work Data on the reliability, calibration standards, and specificity of the assays Help finding the appropriate Novex multiplex assays through our interactive Immunoassay Selection Guide Lists of assays grouped by species and target Our Custom Order Tool for building and ordering your custom Novex multiplex assay
Novex multiplex assay kits offer superior accuracy Protein detection

A
100%
75%
Recovery
50%
25%
0%
FG F
100% 79% 75% 66%
Recovery
50%
25%
To evaluate the Novex multiplex assays, samples of 23 human protein markers were spiked into a sample of human serum, and the sample was processed using the manufacturers instructions for the Novex multiplex assay kit and using similar kits from two other suppliers. The multiplex sample was quantified on the MAGPIX system, and percent recovery was calculated (A) for the individual markers and (B) as an average for the entire group.
-b a 1, sic IL G- 1b CS IL F -1 IL 0 -1 3 IL IL 6 Eo -12 ta xi IL n M -17 IP GM -1 -C a M SF IP M 1b CP IL 1 -1 VE 5 G IF F N IL -g -1 R TN A Fa IL -2 IL IP 7 -1 0 IL -4 IL -8

Novex assay Supplier 3 Supplier 2
17%
0%
Novex assay
Supplier 3
Supplier 2
153
Protein analysis
Novex multiplex assay kits deliver excellent intra-assay precision
A
100%
8% 7%
7%
75%
6% 5%
CV
CV
50%
4% 3%
3%
3%
25%
2% 1%
0%
0%
To evaluate the Novex multiplex assays, samples of 23 human protein markers were spiked into a sample of human serum, and the sample was processed using the manufacturers instructions for the Novex multiplex assay kit and using a similar kits from two other suppliers. The multiplex sample was quantified on the MAGPIX system, and intra-assay precision (CV) was calculated (A) for the individual markers and (B) as an average for the entire group.
Learn more at lifetechnologies.com/luminex.
Protein detection
Novex multiplex magnetic assays
Novex multiplex assays, based on Luminex xMAP (multi-analyte profiling) technology, provide a versatile platform that gives users more flexibility and greater array options for single or multiple analytes. The new line of magnetic multiplex assays was carefully designed and tested to help ensure that sensitivity, range, and correlation are maximized, using the same components as our polystyrene bead Novex multiplex assays. This means that the magnetic and polystyrene assays perform with comperable quality and consistency. Advantages of Novex magnetic multiplex assay kits:
Easier wash steps with the handheld 96-well magnetic separator Enables automation of wash steps Eliminates the hassle of vacuum manifold washes and clogging of filter plates
Learn more at lifetechnologies.com/luminexassays.
154
FG Fba 1, sic IL G- 1b CS IL F -1 IL 0 -1 3 IL -6 IL Eo -12 ta xi IL n M -17 IP GM -1 -C a M SF IP M 1b CP IL 1 -1 VE 5 G IF F N IL -g -1 R TN A Fa IL -2 IL IP 7 -1 0 IL -4 IL -8
Novex assay
Supplier 2
Supplier 3
Novex assay
Supplier 3
Supplier 2
Protein analysis
Luminex 200 MAGPIX
1,000
FLEXMAP 3D
Monkey peripheral blood mononuclear cells (PBMC) were stimulated in vitro with PMA and A2318 for 72 hours. Markers were detected using the Monkey Cytokine Magnetic 28 Plex Panel (Cat. No. LPC0003M), and the results were analyzed on the Luminex 200, MAGPIX, and FLEXMAP 3D Systems.
Luminex instruments for xMAP technology

Learn more about our instruments for evaluating multiplex Luminex assays at lifetechnologies.com/luminexinstrument. Overview of the Luminex instrument systems.
FG ILF- 10 ba si IL c -1 G- b IL CSF -1 0 (2 IL IL ) -1 2p 6 RA 40p N 7 T Eo ES ta xi n IL -1 M 7 IP GM -1a -C M SF IP M 1b CP IL 1 -1 5 EG F IL -5 HG F VE GF IF N -g M DC ITA C M IL IF -1 R TN A Fa IL -2 M IG IL -4 IL -8
Protein detection
Product Feature Optics Hardware Bead compatibility Multiplex capacity Read time for 96-well plate Dynamic range Microtiter plate
MAGPIX System Affordable, efficient, compact size LED/CCD camera Fluorescence imager Magnetic beads only 50 ~60 min 3.5 logs 96-well
Luminex 100/200 System Versatile and efficient; the standard in multiplexing Lasers/APDs/PMTs Flow cytometry based Polystyrene and magnetic beads 100 ~40 min 3.5 logs 96-well
FLEXMAP 3D System High throughput, high plex, automation compatible Lasers/APDs/PMTs Flow cytometry based Polystyrene and magnetic beads 500 ~20 min 4.5 logs 96-well and 384-well
155
Index
Numerical
2D electrophoresis system . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 7-AAD . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Basic Fibroblast Growth Factor (bFGF) . . . . . . . . . . . . . . . . . . .61 BenchMark Protein Ladders . . . . . . . . . . . . . . . . . . . . . . . . 141 BenchPro 4100 Western Processing System . . . . . . . . . . . . 144 BG-11 growth media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 Blasticidin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 BODIPY ceramides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93 Bolt gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136 Brain Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Breast Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 monoclonal antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147 AcTEV Protease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 Actinomycin D . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Advanced reduced-serum media . . . . . . . . . . . . . . . . . . . . . . 41 AKT pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 Alexa Fluor 488 Antibody Labeling Kit . . . . . . . . . . . . . . . . . 106 Alexa Fluor 488 Goat AntiMouse IgG (H+L) . . . . . . . . . . . . 106 Alexa Fluor dyelabeled phalloidins . . . . . . . . . . . . . . . . . . . . 90
Index
ABfinity recombinant
C
Calcein AM probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Cancer research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 Carbenicillin, disodium salt . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 CD Hybridoma Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 CD3/CD28 Dynabeads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79 CD3/CD28/CD137 Dynabeads . . . . . . . . . . . . . . . . . . . . . . . . . 79 Cell counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 Cell culture and cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 Cell depletion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78 Cell dissociation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 Cell Extraction Buffer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 Cell Line Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Cell lysis buffers and protein extraction kits . . . . . . . . . . . . . 133 Cell structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Cell tracing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Cell tracking and neuronal tracing probes . . . . . . . . . . . . . . . .95 Cell viability, proliferation, and health . . . . . . . . . . . . . . . . . . . . 81 CellEvent Caspase-3/7 Green Detection Reagent . . . . . . . . . 85 CellLight reagents Actin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 ER . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93 Golgi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93 Histone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Lysosomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 MAP4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 Mitochondria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Nucleus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Peroxisomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 Tubulin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 CellROX Deep Red Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . 86 CELLstart CTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 CELLstart CTS Substrate . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 CellTrace reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83, 95 CellTracker probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Cellular Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Algae engineering kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 AlgiMatrix 3D cell culture system . . . . . . . . . . . . . . . . . . . . . . 49
Alkaline Phosphatase Live Stain . . . . . . . . . . . . . . . . . . . . . . . . 66 Amino acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 AmnioMAX media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 Ampicillin sodium salt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Analytical and production chromatography . . . . . . . . . . . . . . 131 Antibiotics and antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Antibody and immunoassay selection guides . . . . . . . . . . . . . 37 Antibody Coupling Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130 Antibody Pairs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Antibody purification with protein A/G . . . . . . . . . . . . . . . . . . 131 Antifoaming agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Anti-miR Inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128 Aortic Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 APEX Antibody Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . 107
Assay Buffer Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Astrocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Astrocyte medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 Attune Acoustic Focusing Cytometer . . . . . . . . . . . . . . . 55, 100
B
B-27 Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 BackDrop Background Suppressor . . . . . . . . . . . . . . . . . . . 111 BacMam ion channel targets . . . . . . . . . . . . . . . . . . . . . . . . . . . 88 Bac-N-Blue Baculovirus Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Bacterial selection antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Bac-to-Bac Baculovirus Expression System . . . . . . . . . . . . 117 BaculoDirect Baculovirus Expression System . . . . . . . . . . 117 BaculoDirect Linear DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Baculovirus expression system . . . . . . . . . . . . . . . . . . . . . . . . 117 Balanced salt solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
156
Index
E
Champion Expression Systems . . . . . . . . . . . . . . . . . . . . . . 119 Champion Vector Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 Charcoal-stripped FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Chemically Defined (CD) Hybridoma Media . . . . . . . . . . . . . . . 42 Chemiluminescent ELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . 151 Chemiluminescent substrates for western detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145 Chemokines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47 Click-iT EdU Cell Proliferation Assays . . . . . . . . . . . . . . . . . . 82 Click-iT TUNEL Alexa Fluor Imaging Assay . . . . . . . . . . . . . 85 Co-IP kit for protein complexes . . . . . . . . . . . . . . . . . . . . . . . . 130 Collagenase IV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Concanavalin A Alexa Fluor 594 conjugate . . . . . . . . . . . . . . 93 Coomassie Fluor Orange Protein Gel Stain . . . . . . . . . . . . 142 Corneal Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Countess Automated Cell Counter . . . . . . . . . . . . . . . . . . . . . 53
EKMax Enterokinase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 ELF 97 Endogenous Phosphatase Detection Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 ELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .150 Embryonic stem cellqualified FBS . . . . . . . . . . . . . . . . . . . . . 44 Endoplasmic reticulum and Golgi apparatus . . . . . . . . . . . . . .93 E-PAGE gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Epidermal keratinocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Episomal reprogramming vectors . . . . . . . . . . . . . . . . . . . . . . 64 ER-Tracker dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93 Essential 8 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60 Ethidium homodimer-1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Eukaryotic selection antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . 46 Expi293 Expression System . . . . . . . . . . . . . . . . . . . . . . 42, 116 Express Five SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Extracellular matrices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Index
Cryopreservation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 CSM media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 Custom biology services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33 Custom media for primary cells . . . . . . . . . . . . . . . . . . . . . . . . 54 Custom Novex multiplex assays
F
FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 Fixation and permeabilization for flow cytometry samples . . . . . . . . . . . . . . . . . . . . . . . . . . 105 FLEXMAP 3D System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 FLoid Cell Imaging Station . . . . . . . . . . . . . . . . . . . . 55, 66, 101 Flp-In T-REx System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116 Fluo-4 Direct Calcium Assay Kit . . . . . . . . . . . . . . . . . . . . . . 88 Fluorescence SpectraViewer . . . . . . . . . . . . . . . . . . . . . . . 36, 111 Fluorescent dextran conjugates . . . . . . . . . . . . . . . . . . . . . . . . 95 Fluorescent microspheres . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 FluoroMyelin Green Fluorescent Myelin Stain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 FluxOR Potassium Ion Channel Assay . . . . . . . . . . . . . . . . . 88 FoamAway Irradiated AOF . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 FreeStyle Expression Systems . . . . . . . . . . . . . . . . . . . . 42, 116 FreeStyle MAX Transfection Reagent . . . . . . . . . . . . . . . . . 125
for Luminex technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 Custom primary cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Customer and technical support . . . . . . . . . . . . . . . . . . . . . . . . 37 Cytogenetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 Cytokines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 CytoTune-iPS Sendai Reprogramming Kit . . . . . . . . . . . . . . . 64
D
Dialyzed FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Dispase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Drosophila Expression System (DES) . . . . . . . . . . . . . . . . . . 117 Drosophila S2 Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Dulbeccos Modified Eagle Medium (DMEM) . . . . . . . . . . . . . . 41 Dynabeads cell isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76 Dynabeads for immunoprecipitation . . . . . . . . . . . . . . . . . . . 129 Dynabeads M-270 Epoxy . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Dynabeads M-280 Sheep AntiMouse IgG . . . . . . . . . . . . . . 132 Dynabeads M-280 Sheep AntiRabbit IgG . . . . . . . . . . . . . . 132

G
G-5 Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 Gel-casting accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136 Geltrex LDEV-Free hESC Qualified . . . . . . . . . . . . . . . . . . . . . 62 Geltrex LDEV-Free matrix products . . . . . . . . . . . . . . . . . . . . 49 GeneArt Algae Engineering Kits . . . . . . . . . . . . . . . . . . . . . . 120 GeneArt gene synthesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 GeneArt GeneOptimizer sequence optimization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 Geneticin (G-418) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Dynabeads M-280 Tosylactivated . . . . . . . . . . . . . . . . . . . . . 132 Dynabeads M-450 with secondary antibody against primary antibody . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Dynal CD34 Progenitor Cell Selection System . . . . . . . . . . . 73 DynaMag magnet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
157
Index
Gibco Cell Culture Basics . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Gibco custom media services . . . . . . . . . . . . . . . . . . . . . . . . . 30
Human Microvascular Endothelial Cell, adult dermis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Microvascular Endothelial Cell, neonatal dermis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Neural Stem Cells (H9-Derived) . . . . . . . . . . . . . . . . . 71 Human Pulmonary Artery Endothelial Cell . . . . . . . . . . . . . . . 56 Human Pulmonary Artery Smooth Muscle Cell . . . . . . . . . . . 56 Human Skeletal Myoblasts, large size . . . . . . . . . . . . . . . . . . . 56 Human Skeletal Myoblasts, small size . . . . . . . . . . . . . . . . . . . 56 Human Ultimate ORF Clones . . . . . . . . . . . . . . . . . . . . . . . . 122 Human Umbilical Vein Endothelial Cell . . . . . . . . . . . . . . . . . . 56 Hybridoma-SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Hygromycin B . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
GlutaMAX media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 GoPure Pre-packed
Index
Chromatography Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 Graces Insect Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Growth factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
H
Heart Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Hematopoietic stem cell culture . . . . . . . . . . . . . . . . . . . . . . . . 73 Hematopoietic stem cells (HSCs) . . . . . . . . . . . . . . . . . . . . . . . 73 HEPES buffers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Hibernate media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 High Five cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 High Five Cells (BTI-TN-5B1-4) . . . . . . . . . . . . . . . . . . . . . . . 47 HiMark Protein Standards . . . . . . . . . . . . . . . . . . . . . . . . . . 141 His-tag purification with affinity resins . . . . . . . . . . . . . . . . . . 131 HulaMixer Sample Mixer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 Human Aortic Endothelial Cell . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Aortic Smooth Muscle Cell . . . . . . . . . . . . . . . . . . . . . . 56 Human Astrocyte Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Astrocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human cell isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77 Human Corneal Epithelial Cell . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Coronary Artery Smooth Muscle Cell . . . . . . . . . . . . . 56 Human Dermal Fibroblasts, adult . . . . . . . . . . . . . . . . . . . . . . 56 Human Dermal Fibroblasts, neonatal . . . . . . . . . . . . . . . . . . . 56 Human Epidermal Keratinocyte, adult . . . . . . . . . . . . . . . . . . . 56 Human Epidermal Keratinocyte, adult (animal productfree) . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Epidermal Keratinocyte, neonatal . . . . . . . . . . . . . . . . 56 Human Epidermal Keratinocyte, neonatal (animal productfree) . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Epidermal Keratinocytes, pooled . . . . . . . . . . . . . . . . . 56 Human Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 Human Mammary Epithelial Cell . . . . . . . . . . . . . . . . . . . . . . . 56 Human Melanocyte-adult, lightly pigmented . . . . . . . . . . . . . . 56 Human Melanocyte-neonatal, darkly pigmented . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Human Melanocyte-neonatal, lightly pigmented . . . . . . . . . . 56 Human Melanocyte-neonatal, moderately pigmented . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
I
iBlot 7-Minute Blotting System . . . . . . . . . . . . . . . . . . . . . . . 143 iBlot Western Detection Kits . . . . . . . . . . . . . . . . . . . . . . . . . 145 IEF gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Image-iT Fixation/Permeabilization Kit . . . . . . . . . . . . . . . . 111 Image-iT FX Signal Enhancer . . . . . . . . . . . . . . . . . . . . . . . . 110 Image-iT LIVE Lysosomal and Nuclear Labeling Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 Image-iT LIVE Mitochondrial and Nuclear Labeling Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 iMATCH tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 ImMedia Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 Immunology research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Insect cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Insect expression systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Instruments and accessories . . . . . . . . . . . . . . . . . . . . . . . . . . 99 Invivofectamine 2.0 Reagent . . . . . . . . . . . . . . . . . . . . . . . . . 125 Ion Proton Sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
J
JAK/STAT pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 JC-1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Jump-In Fast Gateway System . . . . . . . . . . . . . . . . . . . . . . 116 Jump-In TI-Gateway System . . . . . . . . . . . . . . . . . . . . . . 116
K
Kanamycin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 KaryoMAX reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 KnockOut media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60, 61
158
Index
L
Labeled secondary antibodies . . . . . . . . . . . . . . . . . . . . . . . . . 109 Labeling and detection products . . . . . . . . . . . . . . . . . . . . . . . 106 Large-scale protein labeling kits . . . . . . . . . . . . . . . . . . . . . . 107 LB medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 LC3B Antibody Kit for Autophagy . . . . . . . . . . . . . . . . . . . . . . . 86 Lectin HPA Alexa Fluor 488 conjugate . . . . . . . . . . . . . . . . . . 93
N
N-2 Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 NativeMark Unstained Protein Standard . . . . . . . . . . . . . . .141 NativePAGE gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Natural Mouse Laminin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 NBD C6-ceramide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93 Negative isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78 Neomycin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Neon Transfection System . . . . . . . . . . . . . . . . . . . . . 55, 65, 124 Neural stem cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 Neural stem cell differentiation . . . . . . . . . . . . . . . . . . . . . . . . 72 Neural stem cells (NSCs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .71 Neuro ELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152 Neurobasal Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48, 71 Neurobiology media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 Neurobiology research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 Neurodegeneration pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Neuronal tracing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 NeuroTrace Nissl stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Novex Semi-Dry Blotter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143 NP-40 Cell Lysis Buffer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 NucBlue Fixed Cell Stain (DAPI special formulation) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 NucBlue Live Cell Stain (Hoechst 33342 special formulation) . . . . . . . . . . . . . . . . . . . 94 Nucleus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 NuPAGE gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
Index
Lentiviral iPSC reprogramming particles . . . . . . . . . . . . . . . . . 64 Lipofectamine reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 LIVE/DEAD Viability/Cytotoxicity Kits . . . . . . . . . . . . . . . . . . . 81 Liver Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 Luminex 100/200 System . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Luminex technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153 LysoSensor dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 Lysosomes and peroxisomes . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 LysoTracker dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
M
M9 minimal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 MagicMedia Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 MAGPIX System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Mammalian cell culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 Mammalian expression systems . . . . . . . . . . . . . . . . . . . . . . . 115 MAPK pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 Mark12 Unstained Standard . . . . . . . . . . . . . . . . . . . . . . . . . 141 MarrowMAX Bone Marrow Medium . . . . . . . . . . . . . . . . . . . . 51 Melanocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Membrane tracers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Mesenchymal stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 MesenPRO RS Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 Microbial culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 Microvascular endothelial cells . . . . . . . . . . . . . . . . . . . . . . . . . 56 Minimal Essential Medium (MEM) . . . . . . . . . . . . . . . . . . . . . . 41 miRNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127 mirVana miRNA Inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . 128 mirVana miRNA Mimics . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128 mirVana PARIS Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 MitoSOX Red Indicator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
O
Opti-MEM reduced-serum medium . . . . . . . . . . . . . . . . . . . . 41 Organelle isolation kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Organelle Lights Nuc-GFP . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Other animal sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
P
PARIS Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .133 pBAD Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 PB-MAX Karyotyping Medium . . . . . . . . . . . . . . . . . . . . . . . . 51 pcDNA vectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116 phosphoELISA kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152 pHrodo dye . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87 PichiaPink Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 PichiaPink Yeast Expression System . . . . . . . . . . . . . . . . . . 118 Plant cell biology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 Pluripotent stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60 Polar tracers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Polymyxin B sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 POROS chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 Positive isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
MitoTracker dyes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .91
Mobile apps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Molecular Probes Handbook . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Monoclonal Antibody Labeling Kits . . . . . . . . . . . . . . . . . . . . . 107 Mouse (C57BL/6) Mesenchymal Stem Cells . . . . . . . . . . . . . . 69 Mouse cell isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77 MSC differentiation kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70 MSC-Qualified FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 Multiplex immunoassays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153 Multiplex magnetic assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154 Mycophenolic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
159
Index
S
Precut blotting membranes . . . . . . . . . . . . . . . . . . . . . . . . . . . 144 Pre-miR Precursors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128 Premo Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83, 86, 88 Primary antibodies for flow cytometry . . . . . . . . . . . . . . . . . . 102 SAIVI Antibody Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . 107 Secondary antibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 Secure FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 SeeBlue Plus2 Pre-stained Standard . . . . . . . . . . . . . . . . . . 141 SeeBlue Pre-stained Standard . . . . . . . . . . . . . . . . . . . . . . . 141 SelectFX Alexa Fluor 488 ER Labeling Kit . . . . . . . . . . . . . . 93 SelectFX Alexa Fluor 488 Peroxisome Labeling Kit . . . . . . . 92 SelectFX Nuclear Labeling Kit, for fixed cells (DAPI, SYTOX Green, 7-AAD, TO-PRO-3 iodide) . . . . . . . . . 94 Selection antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Serum-free media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Sf-900 II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47 Sf-900 III . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Sharp Protein Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Silencer siRNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126 SilverQuest Silver Staining Kit . . . . . . . . . . . . . . . . . . . . . . . 142 SimplyBlue SafeStain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142 Skeletal Muscle Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . 56 Skin primary cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Small-scale protein labeling kits . . . . . . . . . . . . . . . . . . . . . . 107 Sodium bicarbonate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Stealth Select RNAi siRNA . . . . . . . . . . . . . . . . . . . . . . . . . . 126 Stem cell analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67, 70 Stem cell detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66 Stem cell differentiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70 Stem cell reprogramming tools . . . . . . . . . . . . . . . . . . . . . . . . 64 Stem cell research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 Stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60, 69 StemPro Accutase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52, 63 StemPro CD34+ Cell Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 StemPro Differentiation Kits . . . . . . . . . . . . . . . . . . . . . . . . . . 70 StemPro EZPassage Disposable Stem Cell Passaging Tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 StemPro hESC SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60 StemPro Human Adipose-Derived Stem Cell Kit . . . . . . . . . 69 StemPro LipoMAX Supplement . . . . . . . . . . . . . . . . . . . . . . 69 StemPro MSC Serum-Free Medium (SFM) . . . . . . . . . . . . . . 69 StemPro MSC SFM CTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 StemPro MSC SFM XenoFree . . . . . . . . . . . . . . . . . . . . . . . . . 68 StemPro NSC SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 StemPro Rat Alk Phos Expressing Mesenchymal Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 StemPro-34 SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 Streptavidin, Alexa Fluor 488 conjugate . . . . . . . . . . . . . . . . 106
Index
Primary antibodies for western blotting . . . . . . . . . . . . . . . . . 146 Primary Antibody Search Tool . . . . . . . . . . . . . . . . . . . . . . . . . 146 Primary cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Prokaryotic expression systems . . . . . . . . . . . . . . . . . . . . . . . 119 Propidium iodide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Protein A (for IP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130 Protein analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112 Protein detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 Protein expression . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 Protein expression custom services . . . . . . . . . . . . . . . . . . . . 115 Protein expression media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Protein Free Hybridoma Medium-II . . . . . . . . . . . . . . . . . . . . . 42 Protein G (for IP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130 Protein gel electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . 136 Protein quantitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134 Protein sample fractionation . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Protein sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . 129 Protein stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142 Protein standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Protein tag removal with proteases . . . . . . . . . . . . . . . . . . . . 131 Proteins and protein conjugates . . . . . . . . . . . . . . . . . . . . . . . . 95 Protocol Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Pulmonary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 Puromycin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Q
Qdot nanocrystals for multicolor flow cytometry . . . . . . . . 105 Qtracker Cell Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 QuantStudio 12K Flex Real-Time PCR System . . . . . . . . . . . 67 Qubit quantitation system . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
R
Rat Fetal Neural Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 ReadyProbes imaging reagents . . . . . . . . . . . . . . . . . . . . . . 101 Recombinant proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148 Recovery Cell Culture Freezing Medium . . . . . . . . . . . . . . . . 40 Resins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 Rhodamine phalloidin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 RNAi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126 RPMI Medium 1640 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
160
Index
V
Streptomycin sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 SUMO protease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 SYPRO Ruby Protein Gel Stain . . . . . . . . . . . . . . . . . . . . . . . . 142 SYTO 14 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 SYTO 59 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Violet Ratiometric Membrane Asymmetry Probe/Dead Cell Apoptosis Kit . . . . . . . . . . . . . . . . . . . . . . . . 84 ViraPower Lentiviral T-Rex System . . . . . . . . . . . . . . . . . 116 Virtual Cell Stain Tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Vitronectin (VTN-N) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 Voltage sensor probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
Index
SYTO 61 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 SYTO 82 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 SYTO 9 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 SYTOX Dead Cell Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
W
Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 Western blot detection and analysis . . . . . . . . . . . . . . . . . . . . 145 Western transfer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143 WesternBreeze Western Blot Immunodetection Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145 WesternDot 625 Western Blot Kits . . . . . . . . . . . . . . . . . . . . 145
SYTOX Nucleic Acid Stains . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
T
T cell activation/expansion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79 T7 expression systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 Tali Image-Based Cytometer . . . . . . . . . . . . . . . . . . . . . . . 55, 99 TAP growth media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 TaqMan iPSC Sendai Detection Kit . . . . . . . . . . . . . . . . . . . . . 65
X
XCell II Blot Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143 XCell SureLock Mini-Cell . . . . . . . . . . . . . . . . . . . . . . . . . . . 139 XCell4 SureLock Midi-Cell . . . . . . . . . . . . . . . . . . . . . . . . . . 139
TaqMan Protein Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 TC-FlAsH II tetracysteine-based protein detection . . . . . . . 88 TC-ReAsH II tetracysteine-based protein detection . . . . . . . 88 Terrific Broth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 Tissue Extraction Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 TO-PRO-3 Iodide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Transfection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 trc Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 T-REx Inducible Expression System . . . . . . . . . . . . . . . . . . 116 Tris-glycine gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136 TrypLE Express . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 TrypLE Select . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 TrypLE Select CTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Trypsin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 TSA Kit #22, with HRP-streptavidin and Alexa Fluor 488 tyramide . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 TubulinTracker Green . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
Y
Yeast expression system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118 Yeast media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Z
Zenon IgG Labeling Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 Zeocin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 ZOOM IEF Fractionator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Zymogram gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
U
UltraPure 0.5 M EDTA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Umbilical Cord Primary Cells . . . . . . . . . . . . . . . . . . . . . . . . . . 56
161
Life Technologies contacts

UNITED STATES HEADQUARTERS Life Technologies Corp 5791 Van Allen Way Carlsbad, CA 92008 United States Tel: +1 760 602 6500 Toll-Free Tel: 1 800 955 6288 Fax: +1 760 603 7229 Email: customerservices@lifetech.com Email Tech: techsupport@lifetech.com EUROPEAN HEADQUARTERS Life Technologies Ltd Inchinnan Business Park 3 Fountain Drive Paisley, Scotland PA4 9RF United Kingdom Tel: +44 141 814 6100 Toll-Free Tel: 0800 269 210 Toll-Free Tech: 0800 838 380 Fax: +44 141 814 6260 Email: euroinfo@lifetech.com Email Tech: eurotech@lifetech.com Greater China HEADQUARTERS Beijing Office Room 1711 Beijing Silver Tower #2 DongSanHuan North Road Long Sheng Industry Park, Building 203 BDA Beijing, 100027 P.R.C., CHINA Phone: (86) 8008208982 Phone: (86) 4008208982 Email: synsh@lifetech.com Asia Pacific and Japan Headquarters Life Technologies Corp 10 Biopolis Road 4th level, Chromos Singapore 138670 Tel: +65-6861 8638 Email: orders.sg@lifetech.com
Copyright notice
2012 Life Technologies Corporation. All rights reserved. Printed in the USA. CO111234 0912 Life Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies General Terms and Conditions of Sale found on Life Technologies website at www.lifetechnologies.com/termsandconditions. If you have any questions, please contact Life Technologies at www.lifetechnologies.com/support.
Trademarks of Life Technologies Corporation

The trademarks mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners.
Trademarks of other companies

Coomassie is a registered trademark of BASF Aktiengesellschaft. Cy5 and Cy are trademarks or registered trademarks of GE Healthcare. TSA is a registered trademark of PerkinElmer LAS, Inc. Luminex, xMAP, and xPONENT MAGPIX, FLEXMAP 3D, and 100/200 are trademarks or registered trademarks of Luminex Corporation. B-27 is a registered trademark of Southern Illinois University. TaqMan is a registered trademark of Roche Molecular Systems, Inc., used under permission and license. iPhone is a registered trademark of Apple, Inc. CytoTune is a registered trademark of DNAVEC Corporation.

CB Catalog Biología

Transféré par

Informations du document

Description originale:

Titre original

Copyright

Formats disponibles

Partager ce document

Partager ou intégrer le document

Options de partage

Avez-vous trouvé ce document utile ?

Ce contenu est-il inapproprié ?

Droits d'auteur :

Formats disponibles

CB Catalog Biología

Transféré par

Droits d'auteur :

Formats disponibles

2013 Cell biology product and resource guide

Let us help at lifetechnologies.com/support

Its a dynamic time to be working in life sciences.

And we are there to support you at every turn.

Cell biology resources

2013 Life Technologies

Cell biology product and resource guide

Cell culture and cells (Chapter 2) Sample preparation

Cell analysis (Chapter 3)

Classical Gibco media: DMEM, RPMI, and MEM,

Cell biology resources

Cell biology products

Protein analysis (Chapter 4)

Ambion Pre-miR Precursors and Anti-miR Inhibitors,

Cell biology research and pathways

Cell biology resources

Cell culture and cells

CTS brand research use

Class II medical device

Cell biology research and pathways

Cells and culture tools for neurobiology

Neural primary cells:

Primary rat hippocampus neurons

Human neural stem cells

Cell biology research and pathways

Isolated from day-18 Fisher 344 rat embryos

G-5 Supplement Glial cells

Growth of glial cell lines

G-5 Supplement Astrocytes

Rat primary cortical astrocytes

Tools for neural stem cells and progenitor cells.

Neural stem cells

B-27 Supplement, XenoFree N-2 Supplement

KnockOut DMEM/ F-12 Medium

Neural stem cells

Rat fetal neural stem cells

StemPro Neural Supplement

KnockOut DMEM/ F-12 Medium

Neural stem cells

Rat glial precursor cells (rGPCs) Human dopaminergic precursor cells*

StemPro Neural Supplement

* Available as custom order.

Cell biology research and pathways

Neurobasal-A Medium without phenol red Hibernate-A Medium Hibernate -E Medium

Cell biology research and pathways

Neurobasal mediasupport growth of neurons when combined with B-27,

StemPro Accutase Cell Dissociation Reagentdesigned for gentle and efficient

Cell biology research and pathways

Types of cell viability assays.

Cell biology research and pathways

Target gene tranfer plasmid

Clone into baculovirus DNA

Transfect S9 insect cells

Collect BacMam virus

Assay development and screeening

Use BacMam stocks

Transduce mammalian cells

Con gure assays

Perform screen or assay

Cell biology research and pathways

The inflammatory response The AKT pathway The MAPK pathway

The JAK/STAT pathway Neurodegeneration