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Free C-terminus redox-active site Sec and Cys of TrxR were detected by biotin-conjugated iodoacetamide. Different concentrations of AFs were added to NADPH (200 μM) pre-reduced TrxR (0.9 μM) and incubated at 25 °C for 2 h. BIAM was added to label active site free Sec at pH 6.5 and free Cys and Sec at pH 8.5. Results are representative of three independent experiments.
