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Chromotek-RFP-Trap® Booster
Product List
Chromotek-RFP-Trap® BoosterACT-CM-RBOOSTR100 ug
Allele Biotech-Introducing Cost Effectiveness to Research
F
or Research Use Only. Not for Diagnostic or Therapeutic Use.
Purchase does not include or carry any right to resell or transfer this prod-uct either as a stand-alone product or as a component of another product.  Any use of this product other than the permitted use without the express written authorization of Allele Biotech is strictly prohibited
Website: www.allelebiotech.comCall: 1-800-991-RNAi/858-587-6645
(Pacific
Time: 9:00AM~5:00PM)Email: oligo@allelebiotech.com
For Technical Support:Email:FP@allelebiotech.comDescription
RFP-T
rap® coupled to fluorescent dye
 ATTO 594
Specificity
RFP-Trap®
efficiently
 highlights monomeric derivates of dsRed (mRFP1, mCherry,mOrange, mPlum), but not GFP and its variants.- 
Relevance
Red fluorescent proteins (RFPs) are widely used as fluorescent tags.Commonly, a combination of red and green fusion proteins (GFPs) is employed to allow multicolor tracking and analysis of protein-protein interactions. In most applications, the ability to observe fluorescence from red florescent proteins is hindered because the quantum yield and photo-stability of RFPs are usually not sufficient enough for their microscopic detection. This limitation is particularly important for Super Resolution Microscopy applications (e.g. 3D-SIM or STED). Many cell biological methods such as HCl treatment for BrdU-detection, the EdU-Click-iT™ treatment or heat denaturation for FISH, lead to disruption of the RFP signal. By using RFP-Booster_Atto594, a specific RFP-binding protein coupled to the fluorescent dye ATTO 594 (from ATTO-TEC), your RFP signal will significantly increase in stability and brightness.
ATTO 594 is a fluorescent label belonging to the class of Rhodamine dyes.
Fluorescence is excited most efficiently in the range 560 - 615 nm (λabs= 601
nm), a standard 561 nm laser is also suitable for excitation. Maximum fluore-
scence emission falls in the range 615 - 680 nm (λfl= 627 nm).
 
For further information please refer to
http://www.atto-tec.com
Tested Appli-cations
Immunofluorescence
Conc.
1 mg/ml
Storage
Shipped at RT. Store
at 4°C
 Avoid freez
ing
 
1.
Fixation: 4% paraformaldehyde (PFA) or 1:10 formalin (37% formal-dehyde, 10-15% MetOH) in PBS, 10 min., RT.
2.
Wash 3x with PBS containing 0.1% Tween 20 (PBST). Critical: do not let coverslips “dry”.
3.
Permeabilisation: PBS contain-ing 0.5% Triton X-100, 5 min., RT.  Alternatively permeabilise by incu-bating in 100% methanol for 5min at -20°C.
4.
Wash 2x with PBST.
5.
Blocking: 4% BSA in PBST, 10 min, RT.
6.
RFP-Booster incubation: dilute RFP-Booster 1:200-1:400 in blocking buffer and incubate 1 h, RT.
*
7.
Wash 3x 5-10 min in PBST.
8.
 If required counterstain with DNA
fluorescent dyes, e.g. DAPI.
9.
 Before
mounting, briefly rince
 coverslipsbe very rinsed in water to prevent salt crystal formation.
10.
Mount in VectaShield (Vector Labs) or other mounting media with anti-fading agents and seal mounted coverslips with clear nail polish.
Please note: Optimal dilutions/ concentrationsshould be determined by the end user.
*
Tip: RFP-Booster can be added to master-mixduring routine immunostaining, along with the primary or secondary antibodies.
Protocols
HeLa cells expressing mRFP-PCNA fixed and permeabilized. Staining with RFP-Booster_Atto594 (dilution 1/200, 1 h at RT).

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