MMP14 Inhibitor Screening Assay Kit (Colorimetric) (ab139454)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Inhibitor compounds
Overview
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Product name
MMP14 Inhibitor Screening Assay Kit (Colorimetric)
See all MMP14 kits -
Detection method
Colorimetric -
Sample type
Inhibitor compounds -
Assay type
Enzyme activity -
Product overview
Abcam MMP14 Inhibitor Screening Assay Kit (Colorimetric) (ab139454) is a complete assay system designed to screen MMP14 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 (ε=13,600 M-1cm-1 at pH 6.0 and above). The assays are performed in a convenient 96-well microplate format.
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Notes
This kit is useful to screen inhibitors of MMP14, 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.
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Platform
Microplate reader
Properties
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Storage instructions
Please 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 MMP14 Enzyme (Human, Recombinant) 1 x 25µl -
Research areas
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Function
Seems to specifically activate progelatinase A. May thus trigger invasion by tumor cells by activating progelatinase A on the tumor cell surface. May be involved in actin cytoskeleton reorganization by cleaving PTK7. -
Tissue specificity
Expressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors. -
Sequence similarities
Belongs to the peptidase M10A family.
Contains 4 hemopexin-like domains. -
Domain
The 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. -
Post-translational
modificationsThe precursor is cleaved by a furin endopeptidase. -
Cellular localization
Membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Alternative names
- Matrix metallopeptidase 14 (membrane inserted)
- Matrix metalloproteinase 14
- Matrix metalloproteinase-14
see all
Images
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Plot of OD vs. time
Slope=V=2.42E-02 OD/min
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Inhibition of MMP14 by NNGH. Example of inhibitor data. -
Inhibition of MMP14 by NNGH. Example of inhibitor data. -
Example graph for Km and Vmax determination.Km=456 µM
Vmax=42.8 pmol/sec