urgently i want to extract protein for western blot from pancreatic tissue of mouse. for that i m using RIPA buffer and PMSF? whether adding PMSF is sufficient? currently,
PMSF is only available in my lab.
PMSF works to inhibit serine proteases like chymotrypsin, trypsin, etc. but metalloproteases, cysteine proteases, aspartic proteases etc. Try to include Leupeptin as well if you can.
You can replace RIPA buffer with SDS lysis buffer for better outcome.
We use RIPA and PMSF to extract protein from mice pancreas in our lab, so it is fine. However, the RIPA we used has already contain some kinds of proteases inhibitor, like sodium orthovanadate, sodium fluoride, leupeptin. I do not know which kind of RIPA in your lab, so if the RIPA in your lab do not contain such inhibitors, PMSF only is not enough.