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The beads were washed with buffer containing 50 mM Tris pH 8.0, 300 mM NaCl, 12% (v/v) glycerol, 20 mM imidazole and 5 mM 2-mercaptoethanol and eluted with a buffer containing 50 mM Tris pH 8.5, 20 mM NaCl, 300 mM imidazole, 5 mM 2-mercaptoethanol and 1 mM PMSF