Académique Documents
Professionnel Documents
Culture Documents
Blocking Reagents ..................................... 212 - 213 Chemiluminescent Substrates...................218 - 219 Chromatography Buffers ...................................206 Detergents ...........................................................205 Electrophoresis Products ...........................208 - 210 Introduction to ProPure Products ..................196 Labeling Reagents .............................................. 215 ProPure Products ....................................197 - 203 Protease Inhibitors..............................................204 Protein Assays .....................................................207 Protein Transfer................................................... 211 Stripping Buffers.................................................220 Wash Buffers ....................................................... 214 Western Blotting Kits ................................ 216 - 217
Pro-Pure Products
High Purity, Stringently Tested Products for Proteomics Research Free of Protease Activity Functionally Tested Where Applicable
The study of the proteome, or Proteomics, relies on analytical means to identify and quantify proteins in their various forms. It is an extensive field that encompasses techniques in fields such as: Protein Expression and Purification, Protein Separation, Protein Visualization, and Protein Identification and Characterization. As proteins are highly sensitive to a variety of chemicals that can destroy their activity and/or degrade their structure, proteomics grade materials must be used to ensure the integrity and quality of the sample. AMRESCOs Pro-Pure line provides products that are of the highest purity and offer extreme sensitivity. This product line is stringently manufactured and tested to ensure optimal results in proteomics research. By using more sensitive tools, more precise interpretation of the results is possible. Protein Expression and Purification AMRESCOs affinity chromatography buffers are designed for optimal purification and recovery of recombinant proteins. Refer to page 206 of this section. Protein Separation NEXT GEL ready-to-pour acrylamide solutions produce high sensitivity acrylamide gels and enable a high resolution visual comparison between the separated proteins. For a complete listing of all NEXT GEL products as well as AMRESCOs other electrophoresis products, refer to pages 153 - 158 of the Electrophoresis section of this catalog.
Protein Visualization AMRESCO offers a wide range of products for the visualization of proteins both in a gel and membrane bound. For a complete listing and a description of these products, refer to pages 166 - 171 of the Electrophoresis section of this catalog.
Protein Identification AMRESCO offers a wide range of products for the identification of individual proteins. Refer to pages 216 - 220 of this section.
196 PROTEOMICS
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Acr - Bra
Pro-Pure Products
ACRYLAMIDE
Product code Size Price
BENZAMIDINE HYDROCHLORIDE
Product code Size Price
M120
100 g 500 g 1 kg
M141
25 g 50 g 100 g
Proteomics, 204
ACRYL-40
A 40% (W/V) Solution of Ultra Pure Acrylamide. Product code Size Price
M121
100 ml 500 ml
$12.30 $45.40
M225
Electrophoresis, 162
100 g 500 g
$67.40 $244.90
ACRYL/BIS 37.5:1
40% (W/V) SoLutioN A 40% (w/v) solution of Ultra Pure Acrylamide (38.96%) and bis-Acrylamide (1.04%) at a final ratio of 37.5:1. Product code Size Price
BESTATIN
Product code Size Price
M179
Proteomics, 204
10 mg
$125.80
M157
100 ml 500 ml
$17.00 $59.90
ACRYL/BIS 37.5:1
PreMiXed PoWder Ultra Pure Acrylamide and bis-Acrylamide powders blended to achieve a final ratio of 37.5:1. Reconstituted with deionized water, 40 g size has a final volume of 100 ml; 200 g size has a final volume of 500 ml. Reconstitution will produce a 40% (w/v) solution. Product code Size Price
BIS-ACRYLAMIDE
Product code Size Price
M104
50 g 100 g 250 g
M156
40 g 200 g
$28.40 $92.40
BIS-2
2% SoLutioN oF BiS-AcrYLAMide Product code Size Price
AEBSF
(4-(2-AMiNoetHYL)-BeNzeNeSuLFoNYL FLuoride, HYdrocHLoride) Product code Size Price
M193
500 ml
$24.40
M182
50 mg 250 mg 500 mg
Proteomics, 204
M262
Proteomics, 207
500 ml
$35.70
AMMONIUM PERSULFATE
(APS) Product code Size Price
M133
25 g 100 g
$11.20 $26.20
M192
Electrophoresis, 169
1L
$69.40
AMMONIUM SULFATE
Product code Size Price
BORIC ACID
Product code Size Price
M105
1 kg 5 kg
$22.30 $71.40
M139
ANTIPAIN DIHYDROCHLORIDE
Product code Size Price
500 g 1 kg 2.5 kg
M181
Proteomics, 204
5 mg
$41.30
BRADFORD REAGENT
Product code Size Price
M172
Proteomics, 207
Size Price
1L
$108.40
APROTININ
Product code
M167
Proteomics, 204
10 mg 50 mg
$62.90 $257.60
www.amresco-inc.com
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PROTEOMICS 197
Pro-Pure Products
Bra - Gly
M128
M173
Proteomics, 207
1 Kit
$110.40
10 g 25 g 50 g
M125
5g 10 g
$77.00 $135.50
M238
5 Pk
$55.20
BRIj 35
(PoLYoXYetHYLeNe LAurYL etHer) Product code Size
M248
4 pk
$47.70
M106
Proteomics, 205
1 kg 5 kg
$40.00 $139.20
DL-DITHIOTHREITOL
(dtt) Product code Price Size Price
M109
M116
50 g 100 g
$53.10 $100.70
5g 25 g
$69.30 $236.30
E-64
(trans-ePoXYSucciNYL-L-LeucYLAMido(4-GuANidiNo)ButANe) Product code Size Price
CHAPS
(3-[(3-cHoLAMidoProPYL)diMetHYL-AMMoNio]-1-ProPANeSuLFoNAte) Product code Size Price
M186
Proteomics, 204
5 mg
$85.40
M127
5g 10 g 50 g
Proteomics, 205
CHAPSO
(3-[(3-cHoLAMidoProPYL) diMetHYLAMMoNio]-2-HYdroXY-1ProPANeSuLFoNAte) Product code Size Price
M101
500 g 1 kg 2.5 kg
Proteomics, 204
M165
1g 5g 10 g
Proteomics, 205
M134
200 ml 500 ml
$19.70 $25.20
CHYMOSTATIN
Product code Size Price
M184
Proteomics, 204
5 mg
$71.40
M155
100 ml 500 ml
$22.60 $61.00
a-CHYMOTRYPSIN
Product code Size Price
GLYCEROL
Product code Size Price
M102
1g 5g
$52.00 $228.70
M152
1L 4L
$26.00 $113.70
GLYCEROL, STERILE
Product code Size Price
M140
10 g 25 g 50 g
M153
50 ml 100 ml
$13.75 $21.60
198 PROTEOMICS
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Gly - Mop
Pro-Pure Products
M154
100 ml
GLYCINE
Product code Size Price
M264
Proteomics, 206
100 ml 500 ml
$38.50 $191.80
M103
1 kg 5 kg
$32.50 $141.80
IODOACETAMIDE
Product code Size Price
M216
30 g
$73.50
M265
Proteomics, 206
100 ml 500 ml
$27.40 $89.30
M119
10 g 50 g
$58.00 $260.00
GUANIDINE HYDROCHLORIDE
Product code Size Price
LEUPEPTIN
Product code Size Price
M110
M180
Proteomics, 204
5 mg 25 mg
$44.00 $133.90
M113
4L
$736.70
M272
Electrophoresis, 157, Proteomics, 210
1 Kit
$112.20
L-LYSINE MONOHYDRATE
Product code Size Price
M281
Proteomics, 210, Electrophoresis, 158
1 Kit
$291.50
M234
100 g 500 g
$109.50 $263.00
2-MERCAPTOETHANOL
( b-MercAPtoetHANoL) Product code Size Price
M218
Electrophoresis, 162
250 ml 4 X 250 ml
$11.70 $35.00
M131
100 ml 250 ml
$13.40 $18.40
M243
Electrophoresis, 162
250 ml 4 X 250 ml
$55.00 $172.60
M214
Electrophoresis, 162
100 g
$52.00
IMIDAZOLE
Product code Size Price
M136
10 g 50 g 100 g
M215
100 g
$45.50
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PROTEOMICS 199
Pro-Pure Products
Nat - Pep
M203
10 Pk
$18.40
M271
Electrophoresis, 157, Proteomics, 210
1 Kit
$95.40
M158
50 ml 100 ml 500 ml
M254
100 ml 500 ml
$30.80 $104.50
N-OCTYL-b-D-GLUCOPYRANOSIDE
Product code Size Price
M129
500 mg 1g 5g
Proteomics, 205
M255
100 ml 500 ml
$30.80 $104.50
OCTYL-b-D-THIOGLUCOPYRANOSIDE
Product code Size Price
M130
Proteomics, 205
100 mg 1g
$15.90 $36.80
M256
100 ml 500 ml
$30.80 $104.50
M233
Proteomics, 213
5 Pk
$55.20
M257
100 ml 500 ml
$30.80 $104.50
M232
5 Pk
$30.00
Proteomics, 213
M258
100 ml 500 ml
$30.80 $104.50
M245
Proteomics, 213
Price
5 Pk
$20.30
PEPSIN 1:3000
PorciNe Source Product code Size Price
M260
5 ml
$15.90
M142 $78.80
250 g
$43.10
M279
Proteomics, 211
500 ml
PEPSTATIN
Product code Size Price
M183
Proteomics, 204
5 mg 25 mg
$39.00 $102.60
200 PROTEOMICS
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Phe - Sod
Pro-Pure Products
PHENYLMETHYLSUFONYL FLUORIDE
(PMSF) Product code Size Price
M145
Proteomics, 204
5g 25 g
$34.00 $113.80
M307
Proteomics, 204
1 ml
$35.00
1,10-PHENANTHROLINE MONOHYDRATE
(o-Phenanthroline) Product code Size
M266
Electrophoresis, 165
5 ml
$12.00
M135
Proteomics, 204
10 g 50 g
$37.10 $137.80
M252
Electrophoresis, 168
100 ml 1L
$21.80 $73.50
M263
Proteomics, 206
100 ml 500 ml
$27.40 $89.30
PHOSPHORAMIDON
Product code Size Price
M197
500 ml
$47.70
M185
Proteomics, 204
5 mg
$134.90
M227
1 Kit
$139.20
Electrophoresis, 167
SILVER NITRATE
Product code Size Price
M282
Electrophoresis, 171
1L
$81.10
M122
25 g 100 g 500 g
M306
Proteomics, 204
5 ml
$38.20
M138
500 g 1 kg
$20.40 $39.90
M221
Proteomics, 204
1 ml
$35.00
M107
Proteomics, 205
M222
Proteomics, 204
1 ml
$35.00
M250
Proteomics, 204
1 ml
$35.00
M111
Proteomics, 205
100 ml
$20.10
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PROTEOMICS 201
Pro-Pure Products
Sod - Tri
TEMED
(N,N,N',N'-tetrAMetHYLetHYLeNe-diAMiNe) Product code Size Price
M112
Proteomics, 205
200 ml 500 ml
$22.50 $44.00
M146
25 ml 50 ml 100 ml
M137
500 g 1 kg
$24.60 $36.50
M114
1L 4L
$19.30 $68.20
TG BUFFER
PoWder ANd 10X reAdY-PAcK Each pack makes 1L of 10X concentrate. 40L size contains sufficient dry powder to prepare 40L of 1X Buffer. 1X TG solution contains 0.025 M Tris base, 0.192 M Glycine. Product code Size Price
M132
500 g
$15.10
M115
M198
500 ml
$47.70
40 L 2 Pk
$31.80 $34.50
SUCROSE
rNASe ANd dNASe Free Product code Size Price
M117
500 g 1 kg
$17.30 $29.40
M148
1L 4L
$27.30 $76.30
M118
100 ml
$36.20
TG-SDS BUFFER
PoWder ANd 10X reAdY-PAcK Each pack makes 1 L of 10X concentrate. 40 L size contains sufficient dry powder to prepare 40 L of 1X Buffer. 1X solution contains, 0.025 M Tris, 0.192 M Glycine, and 0.1% SDS. Product code Size Price
M149
M231
Proteomics, 213
5 Pk
$60.70
40 L 2 Pk
$36.20 $36.80
THIOUREA
Product code Size Price
M226
100 g 500 g
$45.00 $67.00
M230
Proteomics, 213
5 Pk
$32.00
TRACKING DYE - 2D
Product code Size Price
M217
1 ml
$13.20
Electrophoresis, 165
TRICINE BUFFER
(N-triS(HYdroXYMetHYL)MetHYLGLYciNe) Product code Size Price
M235
Proteomics, 213
5 Pk
$19.90
M159
Electrophoresis, 162
100 g 500 g
$28.60 $93.40
202 PROTEOMICS
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Tri - Zip
Pro-Pure Products
TRITON X-114
Product code Size Price
M151
Electrophoresis, 162
500 g 1 kg
$36.50 $65.00
M144
Proteomics, 205
1L 4L
$23.50 $76.80
TRIS HYDROCHLORIDE
Product code Size Price
TRYPSIN
Product code Size Price
M108
Electrophoresis, 162
500 g 1 kg
$63.60 $85.40
M150
1g 5g
$42.40 $154.20
M191
Proteomics, 204
1g 10 g
$62.40 $442.90
M170
1L
$89.50
M228
5 Pk
$47.70
M171
1 Pk
$79.50
TWEEN 20
(PoLYoXYetHYLeNe-20-SorBitAN MoNoLAurAte) Product code Size Price
M147
50 ml 1L 4L
M168
1L
$89.50
TWEEN 80
(PoLYoXYetHYLeNe-20-SorBitAN MoNooLeAte) Product code Size Price
M126
M169
1 Pk
$86.00
100 ml 1L 4L
UREA
Product code Size Price
M236
5 Pk
$55.20
M123
TRITON X-100
t-octYLPHeNoXYPoLYetHoXYetHANoL Product code Size Price
UREA, 8M SOLUTION
Product code Size Price
M143
Proteomics, 205
1L 4L
$22.40 $72.20
M124
250 ml
$42.13
M277
Electrophoresis, 166
1 Kit
$94.90
M237
5 Pk
$52.50
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PROTEOMICS 203
Protease Inhibitors
Protein Protection
Protease InhIbItors
Extensive Offering Inhibits a Wide Range of Proteases Effectively Preserve the Native State of Proteins Protease Inhibitor Cocktails are Convenient & Easy-to-use
Powerful proteases can begin degrading your protein samples from the moment cells or tissues are lysed. Protein yield and sample quality can be drastically reduced by proteases. Complex protein solutions are usually treated with a mixture of several protease inhibitors in order to preserve the native state of proteins.
Product deScriPtioN Protease inhibitors SPeciFitY oF iNHiBitioN An irreversible serine protease inhibitor of chymotrypsin, trypsin, kallikrein, plasmin and thrombin. Useful as a non-toxic alternative for PMSF. A reversible cysteine and serine protease inhibitor of trypsin, papain and cathepsin A and B. Slightly inhibits plasmin. A competitive and reversible serine protease inhibitor of chymotrypsin, trypsin, kallikrein and plasmin. Not an inhibitor of Factor Xa and thrombin. A protease inhibitor of arginine-selective enzymes like trypsin, kallikrein and thrombin. A competitive inhibitor of aminopeptidases, especially aminopeptidase B, leucine aminopeptidase and tripeptide aminopeptidase. Not an inhibitor of carboxypeptidases. A reversible serine protease inhibitor of a, b, g, d-chymotrypsin. Also inhibits papain and many other cysteine proteases. An irreversible inhibitor of cysteine proteases like papain, calpain and cathepsin B, H, L and S. Also an effective inhibitor of collegenase. A reversible metalloprotease inhibitor useful for chelating metal ions. A reversible inhibitor of cysteine proteases and serine proteases resembling trypsin. Inhibits trypsin, plasmin, papain, kallikrein, thrombin and cathepsin A and B. An aspartic acid protease inhibitor of pepsin, renin and cathepsin D. Also an inhibitor of the HIV protease. Not water soluble. An irreversible serine protease inhibitor of trypsin, chymotrypsin, kallikrein, subtilisin and thrombin. Also inhibits the cysteine protease papain. A metalloprotease inhibitor useful for chelating iron and other divalent metals. A specific inhibitor of thermolysin and other metalloproteases. Also an effective inhibitor of collegenase. Contains a mix of protease inhibitors: AEBSF, Aprotinin, E-64, Bestatin and Leupeptin Contains a mix of protease inhibitors: AEBSF, Aprotinin, E-64, Bestatin and Leupeptin Contains a mix of protease inhibitors: AEBSF, E-64, Bestain, Leupeptin and EDTA Disodium Dihydrate Contains protease inhibitors for mammalian cell use: AEBSF, Aprotinin, E-64, Bestatin, Leupeptin and Pepstatin Contains a mix of protease inhibitors: AEBSF, E-64, Bestain, Pepstatin, Leupeptin and 1, 10-Phenanthroline A serine protease inhibitor specific for trypsin and trypsin-like proteases. WorKiNG rANGe 1.1 1.0 mg/ml (0.4 4 mM) 50 g/ml (74 M) 0.06 2.0 g/ml (0.01 0.3 M) 0.1 0.6 g/ml (0.5 4 mM) 40 g/ml (130 M) 6 60 g/ml (10 100 M) 0.5 10 g/ml (0.14 28.0 M) 1.2 0.5 mg/ml (0.5 1.3 mM) 0.5 g/ml (1 M) 0.7 g/ml (1 M) 17 170 g/ml (0.1 1.0 mM) 4 g/ml (20 mM) 4 330 g/ml (7 570 M) 1X 1X 1X 1X 1X Use at equimolar concentrations code M182-50MG M182-250MG M182-500MG M181-5MG M167-10MG M167-50MG M141-25G M141-50G M141-100G M179-10MG Size 50 mg 250 mg 500 mg 5 mg 10 mg 50 mg 25 g 50 g 100 g 10 mg See PAGe 197
AEBSF* Antipain Dihydrochloride* Aprotinin* Benzamidine Hydrochloride* Bestatin* Chymostatin* E-64* EDTA Disodium Salt* Dihydrate Leupeptin* Pepstatin* Phenylmethylsulfonyl Fluoride (PMSF) 1, 10-Phenanthroline Monohydrate* Phosphoramidon
Protease inhibitor cocktails
197 197
197
197
M184-5MG M186-5MG M101-500G M101-1KG M101-2.5KG M180-5MG M180-25MG M183-5MG M183-25MG M145-5G M145-25G M135-10G M135-50G M185-5MG
5 mg 5 mg 500 g 1 kg 2.5 kg 5 mg 25 mg 5 mg 25 mg 5g 25 g 10 g 50 g 5 mg
199
200 201
201
201
Protease Inhibitor Cocktail, 100X Protease Inhibitor Cocktail, 100X with EDTA Protease Inhibitor Cocktail, Bacterial 100X Protease Inhibitor Cocktail, Mammalian, 100X Protease Inhibitor Cocktail, Plant 100X Trypsin Inhibitor, Soybean
* Product is Animal-free
1 ml 1 ml 5 ml 1 ml 1 ml 1g 10 g
204 PROTEOMICS
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Detergents
Brij 35, 10% Solution Triton X-100, 10% Solution Triton X-114, 10% Solution Tween 20, 10% Solution Detergent Sampler Pack
Contains one 10 ml ampoule of each of the following: Brij 35, 10% Solution Triton X-100, 10% Solution Triton X-114, 10% Solution Tween 20, 10% Solution
Size
1 kg 5 kg 5g 10 g 50 g 1g 5g 10 g 50 ml 100 ml 500 ml 500 mg 1g 5g 100 mg 1g 100 g 250 g 500 g 1 kg 100 ml 200 ml 500 ml 1L 4L 1L 4L 100 ml 1L 4L 100 ml 1L 4L
See PAGe
198 198
Brij-35 CHAPS CHAPSO Nonidet P-40 Substitute N-Octyl-b-D-Glucopyranoside Octyl-b-D-Thioglucopyranoside Sodium Dodecyl Sulfate (SDS)
198
200
200
200 201
Sodium Dodecyl Sulfate (SDS), 10% (w/v) Solution Sodium Dodecyl Sulfate (SDS), 20% (w/v) Solution Triton X-100 Triton X-114 Tween 20 Tween 80
203
Please refer to the Buffers & Detergents section pages 208 - 209 for more information on Detergents
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PROTEOMICS 205
Chromatography
Related Products
Ammonium Acetate Ammonium Phosphate Monobasic Glutathione PBS Potassium Phosphate tris
FuSioN tAG
iMMoBiLized LiGANd
eLutioN BuFFer
Neutral, pH without redox agents. Tag must be accessible in protein structure. High ionic strength, neutral pH
coNceNtrAtioN 5X 5X 5X pH 7.4 7.0 3.0
Product deScriPtioN
Phosphate Chromatography Buffer, Binding Buffer Imidazole Chromatography Buffer, Elution Buffer (His-Tag) Glycine Chromatography Buffer, Elution Buffer (GST Tag)
Acetonitrile Acetonitrile with 0.1% Glacial Acetic Acid Acetonitrile with 0.1% Trifluoroacetic Acid Methanol Methanol with 0.1% Glacial Acetic Acid Methanol with 0.1% Trifluoroacetic Acid Water, Sterile
206 PROTEOMICS
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Protein Assays
Protein Detection
Related Products
NeXt GeL Products Protein Molecular Weight Markers Protein Stain, in-Gel Blue BANdit Protein Stain Silver BuLLit Staining Kit BSA Standards Kit
Product deScriPtioN
code M173-1KIT
Size 1 Kit
M172-1L E531-1.5ML
1L 1.5 ml
197 20
The Biuret Protein Assay is a simple method for determining protein concentration. The amount of protein present in the sample is calculated based on the color change of the copper in the Biuret reagent in the presence of peptide bonds. The intensity of the color change is directly proportional to the number of peptide bonds present.
Product deScriPtioN code M262-500ML Size 500 ml See PAGe 197
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PROTEOMICS 207
Protein Detection
Electrophoresis
Product Summary
NEXT GEL SpriNT NEXT GEL Fluorescent NEXT GEL Fluorescent Sprint NEXT GEL Turbo NEXT GEL 16 x16 cm gels Lp-NEXT GEL HTS-NEXT GEL Native Proteins Native NEXT GEL
Mini-gels
Acrylamide
Denatured Proteins
250 kDa 160 kDa 105 kDa 75 kDa 66.2 kDa 50 kDa 40.0 kDa 97.4 kDa
N XT G L
26.6 kDa 21.5/19.7 kDa
Agarose
A REVOLUTION IN RESOLUTION
Optimized for High Resolution SDS-Page Speed - Pour an Acrylamide Gel in One Minutes High Resolution - Resolve a 14.2 kDa Band from a 14.4 kDa Band on a Mini-gel Broad Range - Separate 3.5 kDa & 212 kDa proteins on same gel Convenience - No Stacking Gel required
35 kDa
30 kDa 25 kDa
For a complete listing and description of NEXT GEL products please refer to pages 153 - 158 of the Electrophoresis section.
*
SdS-PAGe with 10% NeXt GeL. Lane A and d: GE Healthcare Rainbow Molecular Weight Marker. Lane B: 50 g E. coli lysate. Lane c: 100 g E. coli lysate. Lane e: AMRESCO Mid/Low Range Protein Molecular Weight Marker.
Optimized for Running Large Format Gels (16 x 16 cm) in < 3 Hours Quality - Exceptional Resolution at < 1 kDa Convenient - No stacking gel required
Product deScriPtioN MoLecuLAr WeiGHt SePArAtioN rANGe 20-300 kDa code M323-100ML M323-500ML M313-100ML M313-500ML M310-100ML M310-500ML Size 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml See PAGe 130
SdS-PAGe on turbo NeXt GeL. 16 x 16 cm x 1 mm 10% acrylamide gel run at 300 volts for 3 hours. Lane A: GE Healthcare Rainbow Molecular Weight Markers. Lane B: Total E. coli lysates (100 g). Lane c: AMRESCO Mid/Low Range Protein Molecular Weight Marker (Code: J450-200UL).
Includes: NEXT GEL Running Buffer, 20X Each 30 ml will prepare a 16 cm x 16cm x 1.00 mm gel.
10-200 kDa
130
Includes: NEXT GEL Running Buffer, 20X Each 30 ml will prepare a 16cm x 16cm x 1.00 mm gel.
3.5-100 kDa
130
Includes: NEXT GEL Running Buffer, 20X Each 30 ml will prepare a 16 cm x 16 cm x 1.00 mm gel. A B c
*NEXT GEL products are patent pending
208 PROTEOMICS
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Electrophoresis
Protein Detection
*
97.4 kDa 66.2 kDa
Optimized for Running Mini-gels in < 30 Minutes Fast Casting Time - Cast & Polymerize a 10 x 10 x 0.75 cm Mini-gel in < 15 Minutes Convenient - No Stacking Gel required Versatile - Compatible with Standard Staining Procedures & Downstream Applications
PRODUCT DESCRIPTION MoLecuLAr WeiGHt SePArAtioN rANGe 10-200 kDa code M312-100ML M312-500ML M311-100ML M311-500ML Size 100 ml 500 ml 100 ml 500 ml See PAGe 115
40.0 kDa 26.6 kDa 21.5/19.7 kDa 14.4 kDa Silver Stained SDS-PAGE on 10% Sprint Next Gel. Gel was run at 300 V for 30 min. Lane A: AMRESCO Mid/Low Range Protein MW Marker. Lane B: Total E. coli lysate, 50 g.
Includes: NEXT GEL Running Buffer, 20X Each 7.5 ml will prepare a 10 cm x 10 cm x 0.75 mm mini-gel.
3.5-100 kDa
115
Includes: NEXT GEL Running Buffer, 20X Each 7.5 ml will prepare a 10 cm x 10 cm x 0.75 mm mini-gel.
*
A
97.4 kDa 66.2 kDa
Optimized for Immediate Band Visualization Speed - Pour an Acrylamide Gel in One Minute Convenience - Fluorescent Stain incorporated into Gel Solution Enables Immediate Band Visualization Safe - No Need to Prepare or Dispose of Staining & Destaining Solutions
Product deScriPtioN MoLecuLAr WeiGHt SePArAtioN rANGe 10-200 kDa code M290-100ML-KIT M290-500ML-KIT M291-100ML-KIT M291-500ML-KIT Size 100 ml 500 ml 100 ml 500 ml See PAGe 54
40.0 kDa 26.6 kDa 19.7 kDa 14.4 kDa comparison of rapid fluorescent visualization with traditional coomassie staining. A 10% Fluorescent NEXT GEL was run using AMRESCOs Mid/Low Protein Marker (Code: J450-200UL) and NEXT GEL Running Buffer. The gel was run at 175 volts for 1 hour. The protein bands were first visualized by exposure to UV light for 3-5 minutes (A). The same gel (B) was subsequently stained with 0.1% Coomassie R-250 for 3 hours and destained according to standard methods. Fluorescent NEXT GEL offers comparable sensitivity to Coomassie R-250 (100 200 ng/ band) in a fraction of the time.
Includes: NEXT GEL Running Buffer, 20X Each 10 ml will prepare a 10 cm x 10 cm x 0.75 mm mini-gel.
3.5-100 kDa
54
Includes: NEXT GEL Running Buffer, 20X Each 10 ml will prepare a 10 cm x 10 cm x 0.75 mm mini-gel.
Optimized for Fast Mini-gels & Immediate Band Visualization Speed - Cast & Run an SDS-PAGE Mini-gel in Under One Hour Convenience - Fluorescent Stain incorporated into Gel Solution Enables immediate Band Visualization Safe - No Need to Prepare or Dispose of Staining & Destaining Solutions
Product deScriPtioN MoLecuLAr WeiGHt SePArAtioN rANGe 10-200 kDa code M317-KIT-100ML M317-KIT-500ML M318-KIT-100ML M318-KIT-500ML Size 100 ml 500 ml 100 ml 500 ml See PAGe 54
Includes: NEXT GEL Running Buffer, 20X Each 10 ml will prepare a 10 cm x 10 cm x 0.75 mm mini-gel
3.5-100 kDa
54
Includes: NEXT GEL Running Buffer, 20X Each 10 ml will prepare a 10 cm x 10 cm x 0.75 mm mini-gel.
*NEXT GEL products are patent pending
rapid visualization of protein bands on 12.5% Fluorescent Sprint NeXt GeL. Gel was run at 300 V for 30 min., and bands visualized by exposure to UV light for 3 min. Lane A: AMRESCO Wide Range Protein MW Marker. Lane B: Total E. coli lysate. Lane c: Amresco Mid/Low Range Protein MW Marker.
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PROTEOMICS 209
Protein Detection
Electrophoresis
hts N
Optimized for High-Throughput Electrophoresis High-Throughput - Analyze up to 200 Protein Samples in Two Hours Non-Toxic & Biodegradable - Proprietary Agarose Blend Contains No Acrylamide
Product deScriPtioN Includes: HTS Agarose, 60g NEXT GEL Running Buffer, 20X, 2 Packs NEXT GEL Sample Buffer, 4X, 5 ml Contains sufficient reagents for 5 (25 cm x 25 cm) gels (smaller gels can be poured). code M281-KIT Size 1 Kit See PAGe 65, 199
A REVOLUTION IN RESOLUTION
XT G L
MAiN PHoto: HTS NEXT GEL (4%)Stained with Coomassie R-250. iNSet PHoto: HTS NEXT GEL (4%)-Stained with Coomassie R-250. Protein Samples used: MW Standards (Wide) (Code: K494.
lP N
6400 kDa 3200 kDa 1600 kDa 800 kDa 400 kDa 200 kDa
Optimized for Large Protein SDS-PAGE Performance - Resolve 14,000 up to 6 Million Daltons on a Single Gel Versatile - Compatible with Downstream Applications including: Western Blotting, N-terminal Sequencing, Mass Spectometry & MALDI
Product deScriPtioN code M272-KIT Size 1 Kit See PAGe 73, 199
A REVOLUTION IN RESOLUTION
XT G L
Resolution of cross-linked myosin molecule on 1% Agarose HrP prepared with Fluorescent NeXt GeL Buffer. The gel was run at 100 volts for 1 hour. The protein bands were visualized by exposure to UV light for 3-5 minutes. The gel was then stained with 0.1% Coomassie R-250 for 3 hours and destained according to standard procedures (Right). The LP-NEXT GEL Fluorescent procedure offers enormous time savings with sensitivity comparable to Coomassie R-250.
Includes: Agarose HRP, 25 g NEXT GEL Running Buffer, 20X, 500 ml Fluorescent NEXT GEL Buffer, 20X, 125 ml NEXT GEL Sample Loading Buffer, 4X, 5 ml Sufficient materials to run 50 mini-gels.
natIve N
Optimized for Resolution of Both Basic & Acidic Native Proteins Easy-to-use - Non-denaturing Gel Electrophoresis Convenient - Utilizes Standard (Agarose) Electrophoresis Systems Versatile - Compatible with Downstream Applications including: Western Blotting, N-terminal Sequencing, Mass Spectometry & MALDI
A variety of proteins were separated using the Native NeXt GeL electrophoresis Kit. Lane 1: Trypsin Inhibitor. Lane 2: BSA. Lane 3: BSA. Lane 4: Blank. Lane 5: Glucose Oxidase. Lane 6: Glucose Oxidase. (Proteins stained with Coomassie Blue)
A REVOLUTION IN RESOLUTION
XT G L
Product deScriPtioN
code M271-KIT
Size 1 Kit
Includes: Agarose HRP, 25g NEXT GEL Running Buffer, 20X, 500 ml NEXT GEL Sample Buffer, 4X, 5 ml Contains sufficient reagents to perform 30 50 mini gels.
*NEXT GEL products are patent pending
210 PROTEOMICS
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Protein Transfer
Blotting
Protein Detection
The definition of the word detection is most closely linked to the term discovery and discovery is the cornerstone to the life sciences. Although broadly speaking, detection can be viewed as the discovery of anything, in the molecular biology sense, detection largely focuses on the identification of the major biomolecules, DNA, RNA and proteins. Over the years, detection of DNA, RNA and proteins has evolved into not only answering the question of whether a biomolecule is present but specifically, which molecule is present. Commonly referred to as blotting or hybridization, the technique was originally developed in 1975 by British molecular biologist E. M. Southern. The original technique was optimized for the identification of DNA fragments, and appropriately named the Southern Blot. In years to follow, the technique was modified for the detection of RNA, known as the Northern Blot and for protein, known as the Western Blot. With the increasing advances in automation married with the information and software technologies, the birth and growth of microarray technologies has been possible. Based on the traditional blotting methodologies, the microarray and protein array technologies continue to be developed and refined for use as not only research and investigational tools but tools for the fast and accurate diagnosis and predisposition of disease. The fundamental hybridization or blotting technique can be summarized as the very specific interaction of a known fragment of DNA, RNA or antibody with an unknown sample, thereby revealing its identity. Typically the procedures begin with the electrophoretic separation of the molecules, followed by the transfer of these molecules to an immobilized membrane. This membrane is then probed with the known DNA, RNA or antibody, which will bind to the membrane only if the specific target is present on the blot. This presence is revealed through a number of visible modes; including substrate colorimetric reactions, fluorescent molecules, radioactive detection and chemiluminescent substrates.
Related Products
Albumin Blocking Buffers Wash Buffers Horse radish Peroxidase Alkaline Phosphatase Western Blotting Kits
124
transFer buFFers
Product deScriPtioN deScriPtioN Buffer used for the transfer of protein from SDS-PAGE gels to membranes for protein sequencing or Western Blotting Buffer specifically formulated for the transfer of proteins from NEXT GELs to membranes for protein sequencing or Western Blotting coNceNtrAtioN code 10X K872-500ML Size 500 ml See PAGe 34
10X
M279-500ML
500 ml
200
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PROTEOMICS 211
Blotting
Blocking
Technical Tip
Blocking is usually performed using a protein like milk or serum in order to block nonspecific sites on the membrane. Blocking should improve the sensitivity of the assay by reducing the background without interfering with the epitope for antibody binding. Recommendations - PBS interferes with alkaline phosphatase. Therefore, avoid using PBS for blocking when using alkaline phosphatase. - Do not use milk to block membranes when using an avidin-biotin system. Milk contains biotin.
Albumin, Bovine Serum, Fraction V, Heat Shock Isolation Albumin, Bovine Serum, Fraction V, Salt-free Form
20
0903-5G 0903-10G
5g 10 g
20
J642-1ML J642-5ML
1 ml 5 ml
20
K720-50ML K720-500ML
50 ml 500 ml
21
K719-50ML K719-500ML
50 ml 500 ml
21
212 PROTEOMICS
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Blocking
Blotting
blockIng buFFers
High Quality Liquid & Powdered Blocking Buffers Optimize Assay Performance & Improve Sensitivity Ready-To-use Pre-measured Pouches Provide Maximum Convenience
BIO-BLOCK is a blocking buffer optimized for blocking non-specific binding sites on positively charged nylon or PVDF membranes in nucleic acid or protein blotting applications. The resulting lower background increases the signal-to-noise ratio to improve sensitivity and specificity of your blots. BIOBLOCK is available in either 1X PBS or 1X TBS to ensure compatibility with most enzyme systems. Each pack of AMRESCOs Powdered Blocking Buffers will make one liter of 1X solution. These reagents, including TBS with Non-Fat Powdered Milk, TBS with BSA, PBS with Non-Fat Powdered Milk, PBS with BSA and TBS with Tween 20, are designed for use in Western blotting.
Product deScriPtioN code E671-1L E667-1L M232-39.8G-5PK M233-19.8G-5PK M245-10.4G-5PK M230-42G-5PK M231-22G-5PK M235-12.5G-5PK M247-6PK Size 1L 1L 5 pack 5 pack 5 pack 5 pack 5 pack 5 pack 6 pack See PAGe 28 28 200 200 200 202 202 202 29
BIO-BLOCK in 1X PBS BIO-BLOCK in 1X TBS PBS with Non-Fat Powdered Milk PBS with BSA PBS with Tween 20 TBS with Non-Fat Powdered Milk TBS with BSA TBS with Tween 20 Blocking Agent Sampler Pack
Includes 1 of each TBS & PBS pack listed above Excludes Bio-Block & Fish Gelatin Products
Fish gelatin blocking of Western blots - courtesy of dr. eric carlson, case Western reserve university, cleveland, ohio. Whole corneal extracts were immunoprecipitated and resolved by SDS-PAGE on a 10% Fluorescent SPRINT NEXT GEL. Bands were transferred to membrane and blocked with a 5% Fish Gelatin Blocking Buffer in phosphate-buffered saline and Tween 20 (PBST). Primary and secondary antibody incubations were perfomed in PBST prior to development with ECL. The blot was slightly overexposed to show the extremely low background in the control samples (Lanes 1-4) and the experimental samples (Lanes 5-8).
M319-100ML M319-500ML
100 ml 500 ml
53
Wash
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PROTEOMICS 213
Blotting
Peroxide-free detergents
Blot Wash Buffer, 20X PBS Tween 20 Blot Wash Buffer, 20X TBS Tween 20 PBS Powder PBS Tablets PBS, 20X Liquid Concentrate
PBS tablets
PBS, with Tween, pH 7.5 TBS Buffer, 20X Ready-Pack TBS, 20X Liquid Concentrate TBS Buffer Tablets Tween 20, 10% Solution, Peroxide-free 10 ml ampoules
*There is sufficient powder to prepare 10L of a 1X solution by using 9.88 g/L **Each tablet prepares 100 ml of 1X PBS when diluted in water
enzymes
The common use of ELISA, immunoblotting and immunochemical techniques requires the use of high quality enzymes and substrates for the detection of biomolecules. Alkaline phosphatase and horseradish peroxidase are the most commonly employed enzymes for these highly sensitive detection methods. AMRESCO supplies both enzymes and a line of specially prepared colorimetric substrates for each enzyme in powder, tablet and ready-to-use liquid stable forms.
Product NAMe deScriPtioN An enzyme commonly used for the removal of terminal phosphates from the 5 ends of DNA or RNA strands to prevent re-annealing of cohesive ends and facilitate future radiolabeling of the strands. Is most often conjugated to a secondary antibody or cellular component (avidin or biotin). HRP catalyzes the decomposition of hydrogen peroxide in solution causing the oxidation of a number of substrates. Used extensively as a label for immunohistological assays and ELISA. Its exceptional stability in powdered and solution form enhances its desirability as an enzyme conjugate. SPeciFic ActiVitY 3000 U/mg protein code E552-1000UNITS E552-5000UNITS Size 1000 Units 5000 Units See PAGe 21
0343-10000U 0343-25000U
93
93
An enzyme responsible for the degradation of galactosyl b-linkages in glycoproteins, glycolipids and polysaccharides. Can be conjugated to a variety of probes for the chromogenic detection of target sequences using X-Gal.
56
214 PROTEOMICS
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Labeling Reagents
Blotting
D-Biotin
0340-1G 0340-5G
1g 5g
28
Fluorescein Isothiocyanate
0633-500MG
500 mg
53
Rhodamine B Isothiocyanate
0731-100MG 0731-250MG
100 mg 250 mg
105
E497-1MG E497-5MG
1 mg 5 mg
116
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PROTEOMICS 215
Blotting
Chromogenic Kits
K204-1KIT
1 Kit
134
K205-1KIT
1 Kit
134
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Acrylamide Ammonium Persulfate teMed Protein MW Markers ziP reversible Protein Stain
K206-1KIT
1 Kit
134
216 PROTEOMICS
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Chemiluminescent Kits
Blotting
Technical Tip
N220-KIT 1 Kit 134
Weak or low signal Western blots can be caused by proteins that did not transfer properly to the membrane. Precautions: - Use ProAct Membrane Stain (Code: M282-1L) to check that the proteins transferred to the membrane. - After transfer is complete, stain the gel with a total protein stain (Blue BANDit Protein Stain, Code: K2171L) to determine transfer efficiency.
N222-1KIT
1 Kit
134
N223-1KIT
1 Kit
134
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PROTEOMICS 217
Blotting
Chemiluminescent Substrates
VisiGlo Substrates for HRP Offer a Dynamic Range that is Linear for a Longer Period of Time than Other Chemiluminescent Substrates VisiGlo Substrates for AP Feature Long Glow Times of up to 5 Days - Allows for repeated exposure or luminosity reading
visiglo comParIson chart VisiGlo HrP Activation Enzyme Detection Limit Working Solution Emission Duration Stability Recommended Membranes Detection Methods Horseradish Peroxidase (HRP) Picogram Mix 2 components 1-2 hours > 1 year at 4 C Nitrocellulose and PVDF Film VisiGlo PLuS HrP Horseradish Peroxidase (HRP) Femtogram Mix 2 components 4-8 hours > 1 year at 4 C Nitrocellulose and PVDF Film/ Chemiluminescent Imager VisiGlo AP Alkaline Phosphatase (AP) Picogram Ready-to-use > 5 days 2 years at 4 C Nitrocellulose and PVDF Film/ Chemiluminescent Imager VisiGlo PLuS AP Alkaline Phosphatase (AP) Femtogram Ready-to-use > 5 days 2 years at 4 C Nitrocellulose and PVDF Film/ Chemiluminescent Imager
VisiGlo AP Substrate
218 PROTEOMICS
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Chemiluminescent Substrates
Blotting
Comparison of VisiGlo Plus and ECL chemiluminescent substrate sensitivity. Serial dilutions of Mouse IgG (1 ng 31 pg) were separated by SDS-PAGE and transferred to PVDF membranes. Protein detection was performed using HRP-labeled anti-mouse antibody and each respective substrate. Film was exposed for 10 minutes and analyzed for sensitivity and signal to noise ratios. A) VisiGLO PLUS HRP, B) ECL Plus, C) ECL Advance. Sensitivity of VisiGlo HrP. Serial dilutions of mouse (6.1-0.024 pg) IgG were run on SDS-PAGE and transferred to membrane. Blots were developed with VisiGlo HRP and exposed to X-ray film for 10 minutes.
Product deScriPtioN
Size 1 Kit
1 Kit
133
Concentration (pg)
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PROTEOMICS 219
Blotting
Stripping Buffers
Odor-free, Room-temperature Procedure - No hood needed! Convenient Ready-To-use Formulation Minimize Loss of Valuable Antigen
Samarjit Das, Ph.D. Department of Cardiac Pathology Johns Hopkins University School of Medicine, Baltimore, MD
Differential expression pattern of IP3 Receptor Type 1 and Type 3 in Dexamethasone (Dex) treated WEHI 7.2 mouse T cell lymphoma line. Courtesy of Dr. Karen McColl, Case Western Reserve University, Cleveland, Ohio. A single Western blot was used to analyse the expression of IP3 Receptor Type 1 and Type 3 during Dex treatment.. The blot was probed with antibodies to IP3 Recepetor Type 1 and then stripped for 45 minutes with Gentle Review Stripping Buffer prior to reprobing with antibodies to IP3 Receptor Type 3. The different expression pattern obtained with each antibody demonstrates the effectiveness of the stripping and reprobing procedure. type 1: expression increases with Dex treatment.. type 3: expression remains constant with Dex treatment. Actin: Sample loading control.
Product deScriPtioN
code N552-1L
Size 1 liter
See PAGe 58
Related Products
Membranes tris t PBS tween t VisiGlo chemiluminescent Substrates
24 hr Veh 24 hr Veh
Wehi 7.2
24 hr 24 hr
0 hr
differential expression of iP3 type 1 and type 3 receptors in 1 M dexamethasone treated WeHi 7.2 mouse t cell lymphoma line. Courtesy of Dr. Karen McColl, Case Western Reserve University, Cleveland, Ohio. A single Western blot was probed with antibodies to the IP3 receptor Type I and then stripped for 65 minutes with Vantage ReView buffer. After removal of primary and secondary antibodies, the blot was reprobed with IP3 receptor Type 3 antibody. Successful removal of Type 1 antibody is demonstrated by the differential expression of Type 1 and Type 3 antibodies on the same blot.
4 hr 4 hr
8 hr 8 hr
Product deScriPtioN
code
Size 500 ml
N592-500ML
220 PROTEOMICS
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