MMP14 Inhibitor Screening Assay Kit (Colorimetric) (ab139454)

Overview

  • 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.

  • 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.

  • 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
    MMP14 Enzyme (Human, Recombinant) 1 x 25µl
  • 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. Acts as a positive regulator of cell growth and migration via activation of MMP15.
  • 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
    modifications

    The precursor is cleaved by a furin endopeptidase.
  • Cellular localization

    Membrane. Melanosome. Cytoplasm. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Forms a complex with BST2 and localizes to the cytoplasm.
  • Information by UniProt
  • Alternative names

    • Matrix metalloproteinase-14
    • Membrane type 1 metalloprotease
    • Membrane-type matrix metalloproteinase 1
    • Membrane-type-1 matrix metalloproteinase
    • MMP-14
    • MMP-X1
    • Mmp14
    • MMP14_HUMAN
    • MT MMP
    • MT-MMP 1
    • MT1-MMP
    • MT1MMP
    • MTMMP1
    • WNCHRS
    see all

Images

  • Slope=V=2.42E-02 OD/min

  • Km=456 µM

    Vmax=42.8 pmol/sec

Protocols

References

ab139454 has not yet been referenced specifically in any publications.

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