MMP8 Inhibitor Screening Assay Kit (Colorimetric) (ab139452)

Overview

  • Product name

    MMP8 Inhibitor Screening Assay Kit (Colorimetric)
    See all MMP8 kits
  • Detection method

    Colorimetric
  • Sample type

    Inhibitor compounds
  • Assay type

    Enzyme activity
  • Product overview

    Abcam MMP8 Inhibitor Screening Assay Kit (Colorimetric) (ab139452) is a complete assay system designed to screen MMP8 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.

  • Notes

    This kit is useful to screen inhibitors of MMP8, 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.

  • Platform

    Microplate reader

Properties

  • 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
    MMP8 Enzyme (Human, Recombinant) 1 x 22µl
  • Function

    Can degrade fibrillar type I, II, and III collagens.
  • Tissue specificity

    Neutrophils.
  • 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.
  • Cellular localization

    Cytoplasmic granule. Secreted > extracellular space > extracellular matrix. Stored in intracellular granules.
  • Information by UniProt
  • Alternative names

    • Matrix metalloproteinase-8
    • MMP-8
    • Mmp8
    • MMP8_HUMAN
    • Neutrophil collagenase
    • PMNL collagenase
    • PMNL-CL
    see all

Images

  • Slope = V = 1.39E-02 OD/min

  • control slope = 1.39E-02 OD/min

    inhibitor slope (100nM) = 2.73E-03 OD/min

    inhibitor % activity remaining = (2.73E-03/1.39E-02) x 100 = 19.6%

  • Km = 185 µM

    Vmax = 8.90 pmol/sec

Protocols

References

This product has been referenced in:

  • Gonulalan EM  et al. A new perspective on evaluation of medicinal plant biological activities: The correlation between phytomics and matrix metalloproteinases activities of some medicinal plants. Saudi Pharm J 27:446-452 (2019). Read more (PubMed: 30976190) »
See 1 Publication for this product

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