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Cell pellets were thawed in the presence of Lysis Buffer (50 mM Tris pH 8.0, 300 mM NaCl, 10 mM Imidazole, 10% glycerol, 1 mM TCEP, 1 mg/ml lysozyme, (1:1000) DNase I stock for 10 µg/ml final, and 2 protease tablets (SIGMAFAST™ Protease Inhibitor Cocktail Tablets, EDTA-Free Cat# S8830))