ab112140 assay kit quantifies maleimide groups by first reacting a sample with a known amount of thiol present in excess and then assaying the remaining unreacted thiol using 4,4'-DTDP with a molar extinction coefficient of 19,800 M ^-1cm^-1. The amount of maleimide is calculated as the difference between the initial amount of thiol and the amount of unreacted thiol after the complete reaction of all maleimide groups. This spectrophotometric assay for the determination of maleimide groups is a reverse GSH assay. It takes advantage of the high reactivity of thiols of GSH with the maleimide moiety. Maleimide of the sample is allowed to form a stable thiosuccinimidyl linkage with GSH. After the reaction of the sample is complete, the excess GSH, i.e., the remaining thiols of GSH in the reaction mixture, is estimated by using 4,4'-DTDP. The amount of GSH reacted with the sample is titrated to determine the extent of maleimide. For more sensitive maleimide quantitation, we recommend that you use our fluorimetric kit ab112141 that has higher sensitivity.

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