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Expression was induced by IPTG at 20 C, and Glutathione S -transferase (GST) columns (Cytiva) were employed with the Akta Pure System (Cytiva), utilizing buffer A (50 mmol/L HEPES, 500 mmol/L NaCl, 0.5 -mercaptoethanol) and buffer B (50 mmol/L HEPES, 500 mmol/L NaCl, 20 mmol/L reduced glutathione, 0.5 -mercaptoethanol) to purify the GST Tag recombinant protein

Meguid, N

10.1021/acschembio.5b00245 39 DobrianA

Moreover, after the adjustment, the significance was consistent with the previous results