From sample preparation to detection, everything you need for a more efficient and economical western blot is now found in one easy kit. We get our WB results using these reagents, now you can too.
|Cell Lysis Buffer (10X) 9803||15 ml|
|30X Reducing Agent (1.25 M DTT)||1 ml|
|3X Blue Loading Buffer||8 ml|
|Blue Prestained Protein Marker, Broad Range (11-250 kDa) 59329||100 µl|
|BSA 9998||12 g|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl|
|Anti-mouse IgG, HRP-linked Antibody 7076||100 µl|
|PMSF 8553||34.84 mg|
|Tris-Glycine SDS Running Buffer (10X) 4050||300 ml|
|Nitrocellulose Sandwiches 12369||10 Pack|
|Tris-Glycine Transfer Buffer (10X) 12539||300 ml|
|Tris Buffered Saline with Tween® 20 (TBST-10X) 9997||300 ml|
|Nonfat Dry Milk 9999||50 g|
|SignalFire(TM) ECL Reagent A||50 ml|
|SignalFire(TM) ECL Reagent B||50 ml|
All components in this kit are stable for at least 12 months past the reference date indicated on the component label when stored at the recommended temperature and left unused. Upon receipt, BSA and 3X Blue Loading Buffer should be removed from #13005S and stored at 4ºC and at room temperature, respectively. The 30X Reducing Agent and remaining #13005S components should be stored at -20ºC. All #13131S reagents should be stored at room temperature. IMPORTANT: Please refer to the primary antibody datasheet or product webpage for recommended primary antibody dilution buffer and recommended antibody dilution.
NOTE: Please refer to the primary antibody datasheet or product webpage for recommended primary antibody dilution buffer and recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) or smaller membrane; for larger membranes, adjust volumes accordingly.
Proceed to one of the following specific set of steps depending on the primary antibody used.
For Unconjugated Primary Antibodies
For HRP Conjugated Primary Antibodies
For Biotinylated Primary Antibodies
Do not add Anti-biotin, HRP-linked Antibody for detection of biotinylated protein markers. There is no need. The Streptavidin-HRP will also visualize the biotinylated markers.
NOTE: Due to the kinetics of the detection reaction, signal is most intense immediately after incubation.
Protocol Id: 2064
The Western Blotting Application Solutions Kit is designed to conveniently provide reagents needed for western blotting, from cell lysis to protein detection. The reagents in this kit are thoroughly validated with our primary antibodies and will work optimally with the CST western immunoblotting protocol, ensuring accurate and reproducible results. This kit includes sufficient reagents to run 10 mini-gels and complete western blot assays with either rabbit or mouse primary antibodies. All reagents in this kit are available individually.
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