Product Overview : |
The MMP-2 Colorimetric Drug Discovery Kit is a complete assay system designed to screen MMP-2 inhibitors using a thiopeptide as a chromogenic substrate (AcPLG-[2-mercapto-4-methyl-pentanoyl]-LG-OC2H5). The MMP cleavage site peptide bond is replaced by a thioester bond in the thiopeptide. Hydrolysis of this bond by an MMP produces a sulfhydryl group, which reacts with DTNB [5,5’- dithiobis(2-nitrobenzoic acid), Ellman’s reagent] to form 2-nitro-5-thiobenzoic acid, which can be detected by its absorbance at 412 nm (ε= 13,600 M-1cm-1 at pH 6.0 and above). The assays are performed in a convenient 96-well microplate format. The kit is useful to screen inhibitors of MMP-2, a potential therapeutic target. An inhibitor, NNGH12, is also included as a prototypic control inhibitor. Thiol inhibitors should not be used with this kit, as they may interfere with the colorimetric assay. |
Storage : |
Store all components except the microplate (room temperature) at -70°C for the highest stability. The MMP-2 enzyme should be handled carefully in order to retain maximal enzymatic activity. It is stable, in diluted or concentrated form, for several hours on ice. As supplied, MMP-2 enzyme is stable for at least 5 freeze/thaw cycles. To minimize the number of freeze/thaw cycles, aliquot the MMP-2 into separate tubes and store at -70°C. When setting up the assay, do not maintain diluted components at reaction temperature (e.g. 37°C) for an extended period of time prior to running the assay. |