CellMosaic’s Thiol Assay Kit with Purification is designed to assay the free thiol groups of a biopolymer after reducing or labeling with small molecule thiol-modification reagents. CellMosaic routinely uses this kit for its internal bioconjugation-related research.
This product can be used together with our antibody or other biopolymer labeling and conjugation kits to optimize loading. Examples of these kits include:
- Antibody MMAE Conjugation Kit (With VC-PAB Linkage) (Cat#: CM11409, CM11409x3)
- Antibody Mc-MMAF Conjugation Kit (CM11422 and CM11422x3)
- Antibody MMAF Conjugation Kit (with VC-PAB Linker)(CM11425 and CM11425x3)
- F(ab')2 MMAE Conjugation Kit (With VC-PAB Linkage) (CM11416 and CM11416x3)
Please follow the protocols for using the thiol assay kit along with these conjugation kits.
About the Assay:
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.
Application of the product:
- Purification of a small amount of biopolymer after a reducing or thiol labeling reaction
Assay the free thiol groups
Key Features of the product:
Less than 30 minutes of preparation and assay time. Fast and easy to use.
- Each package is sufficient for 10 purifications and 5 assays (100 μL per assay volume).