Modified RIPA Buffer
Recipe
10 mM Tris, pH 7.4
100 mM NaCl
1 mM EDTA
1 mM EGTA
1 mM NaF
20 mM Na4P2O7
2 mM Na3VO4
1% Triton X-100
10% glycerol
0.1% SDS
0.5% deoxycholate
plus protease inhibitors: Aprotinin, leupeptin, pepstatin: 1ug/ml each
Add 1mM PMSF immediately before use.