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A 12.5 g cell pellet was resuspended in 40 ml buffer B (25 mM Tris-HCl, pH 7.5, 10% glycerol, 0.01% Igepal CA-630, 2 mM 2-Mercaptoethanol, 5 mM Imidazole) containing 500 mM KCl and a cocktail of protease inhibitors (PIs) (2 g/ml each of aprotinin, chymostatin, leupeptin, and pepstatin A) and lysed by sonication (QSonica Q700) at an amplitude of 40 for 2.5 min with 20 s pulses