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).
產(chǎn)品描述
CellMosaic的巰基測定試劑盒旨在測定生物聚合物的游離巰基含量,例如部分還原的抗體。CellMosaic經(jīng)常使用該試劑盒進行內(nèi)部生物綴合相關(guān)研究。該測定基于Ellman的測定,使用5 '5'-二硫代-雙-(2-硝基苯甲酸)(DTNB)(Ellman,G. L. 1959年,組織巰基。生物化學(xué)與生物物理學(xué)導(dǎo)論82,70-77)。硫醇與DTNB反應(yīng)生成2-硝基-5-硫代苯甲酸(TNB)。TNB為橙子,在412 nm處的消光系數(shù)為14,150 M-1cm-1(Riddles,P.W.布拉沃河L.,和Zerner,B. 1983年,Ellman試劑的重新評估。Methods Enzymol. 91,49-60)。硫醇基的量將根據(jù)產(chǎn)生的TNB的量計算。每個包裝四個微量離心管。每個包裝足以進行10次測定(每個測定體積100 μL)。