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After loading the supernatant, the column was washed with 20 column volumes of 90% buffer A (0.5 M NaCl, 0.2 M sodium phosphate, and 20 mM imidazole) and 10% buffer B (0.5 M NaCl, 0.2 M sodium phosphate, and 0.5 M imidazole) followed by elution with 100% buffer B
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