|Cytoplasmic Isolation Buffer (CIB)||10 ml|
|Membrane Isolation Buffer (MIB)||10 ml|
|Cytoskeletal/Nuclear Isolation Buffer (CyNIB)||5 ml|
|Protease Inhibitor Cocktail (100X) 5871||250 µl|
Store at -20°C. Upon receipt, Protease Inhibitor Cocktail 5871 should be stored at 4°C. All other components should be stored at -20°C.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
For adherent cells
For both adherent and suspension cells
Table 1: Volumes in μl for WCL or buffer at indicated cell numbers.
|2.5 x 106 cells||5 x 106 cells||7.5 x 106 cells||1 x 107 cells|
Protocol Id: 584
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