PME Dynein Buffer
0.1 m PIPES (pH 6.9)
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An alternative to PIPES (pKa = 6.8) is MES (pKa = 6.1), another sulfonic acid–based buffer.
2 mm EGTA
1 mm MgSO4
1 mm DTT (or DTE)
1 mm GTP
It may be of value to include one or more of the following inhibitors to reduce endogenous protease activity in the extracts being studied: 2 mm 4-benzenesulfonyl fluoride hydrochloride (AEBSF or Pefabloc), 1 mm PMSF, Halt Protease Inhibitor cocktail, or a mixture of 1 mm PMSF, 20 µg/mL leupeptin, 10 µg/mL antipain, and 1 µm pepstatin A.
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