Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Inhibitor compounds
Product nameMMP12 Inhibitor Screening Assay Kit (Colorimetric)
See all MMP12 kits
Sample typeInhibitor compounds
Assay typeEnzyme activity
Abcam's MMP12 Inhibitor Screening Assay Kit (Colorimetric) (ab139441) is a complete assay system designed to screen MMP12 inhibitors using a thiopeptide as a chromogenic substrate (Ac-PLG-[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 (e=13,600 M-1cm-1 at pH 6.0 and above). The assays are performed in a convenient 96-well microplate format.
This kit is useful to screen inhibitors of MMP12, a potential therapeutic target. The MMP inhibitor NNGH 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 instructionsPlease refer to protocols.
Components 1 x 96 tests 96-well Clear Microplate (1/2 Volume) 1 unit Colorimetric Assay Buffer 1 x 20ml MMP Inhibitor 1 x 50µl MMP Substrate 1 x 50µl MMP12 Enzyme (Human, Recombinant) 1 x 14µl
FunctionMay be involved in tissue injury and remodeling. Has significant elastolytic activity. Can accept large and small amino acids at the P1' site, but has a preference for leucine. Aromatic or hydrophobic residues are preferred at the P1 site, with small hydrophobic residues (preferably alanine) occupying P3.
Tissue specificityFound in alveolar macrophages but not in peripheral blood monocytes.
Sequence similaritiesBelongs to the peptidase M10A family.
Contains 4 hemopexin-like domains.
DomainThe conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
Cellular localizationSecreted > extracellular space > extracellular matrix.
- Information by UniProt
- EC 188.8.131.52
- Macrophage elastase
ab139441 has not yet been referenced specifically in any publications.