Estimation of Disulfide Bonds Using Ellman’s Reagent
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Ellman’s reagent 5,5′-dithiobis(2-nitrobenzoic acid)(DTNB) was first introduced in 1959 for the estimation of free thiol groups (1). The procedure is based on the reaction of the thiol with DTNB to give the mixed disulfide and 2-nitro-5-thiobenzoic acid (TNB) which is quantified by the absorbance of the anion (TNB2−) at 412 nm.
KeywordsDisulfide Bond Reaction Buffer Free Thiol Disulfide Thiol Mixed Disulfide