M2 lysis buffer

50 mM Tris-base (pH 7.4)

150 mM NaCl

10% glycerol

1% Triton X-100

0.5 mM EDTA

0.5 mM EGTA

50 mM NaF

40 mM β-glycerophosphate

5 mM tetrasodium pyrophosphate

0.1 mM sodium vanadate

10 μg/ml aprotinin

5 μg/ml leupeptin

2 mM phenylmethylsulfonyl fluoride (PMSF), fresh

Prepare immediately before use and chill (ice-cold) before use.