CellMosaic’s thiol assay kit is designed to assay the free thiol group content of a biopolymer, such as a partially reduced antibody. CellMosaic routinely uses this kit for its internal bioconjugation-related research.
The assay is based on Ellman’s assay using 5’5’-dithio-bis-(2-nitrobenzoic acid) (DTNB) (Ellman, G.L. 1959, Tissue sulfhydryl groups. Arch Biochem Biophys. 82, 70–77). Thiol is reacted with DTNB to generate 2-nitro-5-thiobenzoic acid (TNB). TNB is orange in color and has an extinction coefficient of 14,150 M-1cm-1 at 412 nm (Riddles, P.W. Blakeley, R. L., and Zerner, B. 1983, Reassessment of Ellman’s reagent. Methods Enzymol. 91, 49-60). The amount of thiol groups will be calculated based on the amount of TNB generated.
Four micro-centrifuge tubes per package. Each package is sufficient for 10 assays (100 μL per assay volume).
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CellMosaic的巯基测定试剂盒旨在测定生物聚合物的游离巯基含量,例如部分还原的抗体。CellMosaic经常使用该试剂盒进行内部生物缀合相关研究。该测定基于Ellman的测定,使用5 '5'-二硫代-双-(2-硝基苯甲酸)(DTNB)(Ellman,G. L. 1959年,组织巯基。生物化学与生物物理学导论82,70-77)。硫醇与DTNB反应生成2-硝基-5-硫代苯甲酸(TNB)。TNB为橙子,在412 nm处的消光系数为14,150 M-1cm-1(Riddles,P.W.布拉沃河L.,和Zerner,B. 1983年,Ellman试剂的重新评估。Methods Enzymol. 91,49-60)。硫醇基的量将根据产生的TNB的量计算。每个包装四个微量离心管。每个包装足以进行10次测定(每个测定体积100 μL)。