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Briefly, a 1 ml reaction mix (500 nM TGR, 100 M NADPH, and 2050 M inhibitor) was incubated at room temperature for 30 min for fast inhibitors, or 2 h for slow inhibitors

Buffer Exchange: Ensure the protein is in an amine-free buffer (e.g., PBS, 100 mM sodium phosphate, 150 mM NaCl) at pH 7.2-7.5

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In contrast, CST IV is a diverse group distinguished by a lower presence of Lactobacilli , typically with the highest pH median