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Development and validation of GluICAT methodology. A, Schematic illustrating the GluICAT strategy of differential tagging of reduced and glutathionylated thiols. Reduced thiols are tagged with light (12C) ICAT, whereas glutathionylated thiols are reduced by Grx1 then tagged with heavy (13C) ICAT, and subsequently detected by LC/MS/MS. B, H:L ratios for GAPDH (untreated) and GAPDH-SG (with (+) or without (−) Grx1) from the tryptic GAPDH peptide VPTPNVSVVDLTCR (mean of 3 replicates).
