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4-Vinylpyridine is used as a derivatizing reagent for free thiols such as GSH. It is used in the Glutathione colorimetric assay to remove reduced GSH, so that the oxidized Glutathione (GSSG) concentration can be measured. 4-Vinylpyridine is a better derivatizing reagent for GSH when compared to N-ethyl-maleimide (NEM) as N-ethyl-maleimide is a potent inhibitor of glutathione reductase. The treatment of samples with 4-vinylpyridine removes all the free thiols present in the sample leaving only GSSG which can be quantified in the same way as total glutathione using Ellman’s Reagent.
4-Vinylpyridine alkylates cysteine and cystine residues (after reduction) in proteins to give derivatives that are stable to acid hydrolysis and so it is used in analysis of proteins2. Its alkylating property also enables it to be used for preparation of proteins from PAGE for peptide mapping by MALDI-MS and MALDI-TOF.
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