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A 10 gram pellet of cells was resuspended in 30 ml buffer T (25 mM Tris-HCl, pH 7.5, 10% glycerol, 0.5 mM EDTA, 1 mM DTT, 0.01% Igepal CA-630) containing 500 mM KCl and a cocktail of protease inhibitors (PIs) (5 μg/ml each of aprotinin, chymostatin, leupeptin, and pepstatin A) and lysed with a Dounce homogenizer

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