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==Protocol==
samples are on ice/in cold * Lyse Scrape cells in lysis buffer, such as RIPA buffer with protease inhibitors and phosphatase inhibitors (see [[RIPA Buffer]]).
* For each well in a 12 well plate lyse in 1 ml.
* rotate combine 0.5 ml lysate with , 2 ul of antibody and 25 ul of protein A/G plus agarose beads (catalog # sc 2003 santa cruz).* rotate 1 hour-overnight in cold room* spin 2 minutes at 5000 rpm and aspirate supernatent, carefully avoiding beads* wash X3: add 1 ml lysis buffer, mix by inverting tube, spin and aspirate supernatent as before.* spin once more and aspirate all supernatant.* add 50 ul of sample buffer, boil 5 minutes* to get rid of beads - open cap, prick bottom with 27G needle (up to halfway of bevel). can
