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Free Thiol Assay Kit

Components:

Thiol Assay Solution: 20 ml; store at 4°C (200 assays)
10x Cell Lysis Solution: 25 ml; store at 4°C

Product Description: The detection and measurement of free thiols (sulfhydryl groups) associated with small molecules (cysteine, glutathione, coenzyme A and short peptides) and cysteine residues on proteins provides important information regarding the redox status and metabolic flux of the biological system. Oxidative stress profoundly impacts cell function both by direct and indirect oxidation of sulfhydryl groups. The kit is based on quantification of free thiol groups by Ellman's reagent (5,5'-dithiobis-(2-nitrobenzoic acid); DTNB), which releases
2-nitro-5-thiobenzoate (NTB) upon reaction with free thiol groups. NTB exhibits an intense yellow color that can be quantified by spectrophotometer at 410 - 415 nm using an extinction coefficient of 14,150/M-cm. The assay, performed at room temperature, can be completed in 5 min. As low as 2 - 10 uM free thiols can be detected, including low-molecular weight thiols and those thiol groups on proteins. Each kit provides sufficient material for 250 assays, and includes a cell/tissue extraction solution for preparation of crude lysates for the assay. The kit is for research and diagnostic applications.