Protease inhibitors used in CHIP - (Feb/20/2007 )
Hi, I will do CHIP. It is said the protease inhibitors are PMSF, aprotinin and pepstain in the protocol. Someone used PMSF plus cocktail inhibotr when wahsing the dishes, and only cocktail when lysising the cells. I wonder if I can use Complete Cocktail inhibitor only from Roche? I really appreciate your help, thanks.
I am not sure what is in the cocktail, but PMSF is an irreversible inhibitor that has a short half life when used. I would use this from a stock. In the end protease inhibitors are cheap, repeating an experiment is not. IMHO, I would hit with PMSF, aprotinin, pepstatin, soybean trypsin inhibitor (STI), and leupeptinin. Don't be afraid to add a little more than needed, degradation is bad.
we use proteinase inhibitor cocktail from roche for ChIP anaysis.
works a treat, but is more expensive than a homebrew equivalent.