IP from frozen protein extracts - (Aug/11/2006 )
sorry I forgot the other questions.
With the cells I used (fibroblast, and fibroblast-like), 0.1 % was enough.
I add the lysis buffer, mix by pipetting, and let incubate on ice 20 min at least.
Then I centrifuge at 14000g 15 min, and freeze the supernatant at -80°C.