I am new to western bloting, i need to isolate protein ( bcl2/ bax/ nf kappa b/ caspase) by western blotting from breast cancer cells, i will use ripa buffer with protease inhibitor cocktail , do i need to add sodium orthovanadate ???
I had excellent results with RIPA and protease inhibitor cocktail with breast cancer cells. I used to add phosphatase inhibitors because I wanted to study phosphorylation. The only thing I remember is that we had RIPA aliquoted and the protease and phosphatase inhibitors were added prior to use not to aliquots.