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The supernatant was re-equilibrated by adding 25% volume of re-equilibration buffer (150 mM HEPES pH 8.0, 700 mM KOAc, 15 mM Mg(OAc) 2 , 42% (v/v) glycerol, 5 mM NDSB-256 and 0.5 mM TCEP, with 1 tablet of cOmplete Mini EDTA-free Protease Inhibitor Cocktail per 10 ml of buffer), then further clarified by centrifugation at 60,000 g for 20 min at 10 °C
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