Washing buffer (store at 4°C for up to 6 months)

Reagent	Final concentration	Amount
Tris-HCl	50 mM	6 g
NP40	1%	10 mL
Sodium dodecyl sulfate	0.1% (w/v)	1 g
Sodium chloride	150 mM	8.77 g
Sodium deoxycholate	0.5% (w/v)	2.5 g
Sodium ascorbate	10 mM	1.98 g
EDTA	1 mM	0.29 g
HALT protease inhibitor (add upon use)	1×	10 mL (100×)
Trolox	5 mM	1.25 g
Sodium azide	10 mM	0.65 g
PBS	/	Fill up to 1000 mL