HPLC-based assays for enzymes of glutathione biosynthesis.

Glutamate cysteine ligase and glutathione synthase carry out the two-step synthesis of glutathione. The fluorescent thiol-reactive compound monobromobimane is used to derivatize reaction products in an HPLC-based assay with fluorescence detection. The assay described in this unit can be adapted for tissue homogenates or cultured cells.