EZCell™ Thiol Detection Kit

Thiols are organosulfur compounds that contain a free sulfhydryl group (-SH). The most predominant intracellular thiols in eukaryotes are cysteine, glutathione and cysteine-containing proteins. Thiols are vital in mediating cellular redox chemistry, preventing oxidative damage by reactive oxygen species and assisting in proper protein folding via the formation of disulfide bridges. Under physiological conditions, glutathione (GSH) is the most abundant intracellular thiol. High levels of reduced glutathione in the cytosol help maintain a reduced redox state, as GSH can react with the electrophilic xenobiotics and reduce protein thiols to support a variety of enzymatic redox reactions. Depletion of intracellular GSH and other thiols results in the buildup of lipid peroxidation products, indiscriminate oxidation of protein cysteine residues and is a strong activator of apoptosis. BioVision’s EZCell™ Thiol Detection Kit utilizes a cell-permeable, thiol reactive fluorogenic Thiol Dye. The dye is strongly quenched and is essentially non-fluorescent until it reacts with an intracellular thiol moiety to generate a bright green fluorescence. The fluorescence can be quantified by measuring the fluorescence intensity in the FL1 channel (Ex/Em = 492/517 nm) of a flow cytometer. The thiol dye is selective for free reduced thiols, reacting with reduced GSH and cysteine to form highly fluorescent adducts, and does not label oxidized GSSG. The fluorescent conjugates are stable and are directly proportional to the intracellular thiol level. The EZCell™ Thiol Detection Kit provides a rapid, non-radioactive and sensitive method for measuring the overall intracellular thiol level in living cells and for studying the effects of oxidative stress and other thiol-modifying conditions.