Protease Inhibitor Cocktail

4 mL 2× NPB

2.4 mL 10% bovine serum albumin (BSA; e.g., Sigma-Aldrich A3294; 10% w/v with pH adjusted to 7.6 with KOH, aliquoted, and stored at −20°C)

8 µL leupeptin (e.g., Sigma-Aldrich 11017128001; 10 mg/mL in DMSO that has been stored at −20°C)

8 µL 0.3 m phenylmethylsulfonyl fluoride (PMSF; e.g., Sigma-Aldrich 78830; that has been aliquoted and stored at −20°C)

1.6 mL sterile Nanopure H₂O

Freshly prepare 8 mL and store on ice. Thaw leupeptin and PMSF stocks in a 37°C water bath just before use (vortex in case crystals are formed). Use gloves when handling protease inhibitors.

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