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2, step 3), and the pellet was saved for further analyses Protein refolding The 5 ml of solubilized recombinant protein was added to 300 ml of refolding buffer containing 50 mM Tris-HCl, pH 8.5, 0.4 M sucrose, 10% glycerol, 0.5% Triton X-100, 0.3 mM glutathione disulfide (GSSG), and 3 mM glutathione (GSH), at a rate of 1 ml/hour and stirred at 200 rpm at 4 °C

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