Wash buffer F

0.2 mM EDTA

15% (v/v) glycerol

2 mM MgCl₂

150 mM NaCl

0.01% (v/v) Nonidet P-40

20 mM Tris-Cl (pH 7.9)

Store solution for up to 24 h at 4°C.

Immediately before use, add:

10 mM β-glycerophosphate

1 μg/mL aprotinin

0.5 mM benzamidine hydrochloride

1 mM DTT (dithiothreitol)

2 μg/mL leupeptin

0.2 mM phenylmethylsulfonyl fluoride