|Cell Lysis Buffer (10X) 9803
|30X Reducing Agent (1.25 M DTT)
|3X Blue Loading Buffer
|Blue Prestained Protein Marker, Broad Range (11-250 kDa) 59329
|Anti-rabbit IgG, HRP-linked Antibody 7074
|Anti-mouse IgG, HRP-linked Antibody 7076
|Tris-Glycine SDS Running Buffer (10X) 4050
|Nitrocellulose Sandwiches 12369
|Tris-Glycine Transfer Buffer (10X) 12539
|Tris Buffered Saline with Tween® 20 (TBST-10X) 9997
|Nonfat Dry Milk 9999
|SignalFire(TM) ECL Reagent A
|SignalFire(TM) ECL Reagent B
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
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