MMP3 Inhibitor Screening Assay Kit (Colorimetric) (ab139439)
- Datasheet
- References
- Protocols
Overview
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Product nameMMP3 Inhibitor Screening Assay Kit (Colorimetric)
See all MMP3 kits -
Detection methodColorimetric
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Sample typeInhibitor compounds
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Assay typeEnzyme activity
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Product overview
Abcam's MMP3 Inhibitor Screening Assay Kit (Colorimetric) (ab139439) is a complete assay system designed to screen MMP3 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 MMP3, 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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PlatformMicroplate reader
Properties
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Storage instructionsPlease refer to protocols.
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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 MMP3 Enzyme (Human, Recombinant) 1 x 30µl -
Research areas
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FunctionCan degrade fibronectin, laminin, gelatins of type I, III, IV, and V; collagens III, IV, X, and IX, and cartilage proteoglycans. Activates procollagenase.
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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.
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Cellular localizationSecreted > extracellular space > extracellular matrix.
- Information by UniProt
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Alternative names
- CHDS6
- Matrix metalloproteinase 3
- Matrix metalloproteinase-3
see all
Images
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Plot of OD vs. timeSlope = V= 1.185E-02 OD/min
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Inhibition of MMP3 by NNGH (1.3µM)control slope = 1.4E-02 OD/min
inhibitor slope = 1.3E-04 OD/min
inhibitor % activity remaining = (1.3E-04/1.4E-02) x 100 = 0.9%
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Inhibiton of MMP3 by NNGH -
Example graph for Km and Vmax determinationKm = 276 µM
Vmax = 10.6 pmol/sec
Protocols
References
ab139439 has not yet been referenced specifically in any publications.
