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Subsequently, 20 l of the standard GSH or lysates were mixed with 250 l of 0.1 M phosphate buffer pH 7.6, and 50 l of 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB) (cat# 69-78-3 Sigma-Aldrich, USA), then kept in the dark for 5 min