MMP13 Inhibitor Screening Assay Kit (Colorimetric) (ab139450)

Overview

  • Product name

    MMP13 Inhibitor Screening Assay Kit (Colorimetric)
    See all MMP13 kits
  • Detection method

    Colorimetric
  • Sample type

    Inhibitor compounds
  • Assay type

    Enzyme activity
  • Product overview

    Abcam MMP13 Inhibitor Screening Assay Kit (Colorimetric) (ab139450) is a complete assay system designed to screen MMP13 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-1 cm-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 MMP13, 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
    MMP13 Enzyme (Human, Recombinant) 1 x 53µl
  • Function

    Degrades collagen type I. Does not act on gelatin or casein. Could have a role in tumoral process.
  • Tissue specificity

    Seems to be specific to breast carcinomas.
  • Involvement in disease

    Defects in MMP13 are the cause of spondyloepimetaphyseal dysplasia Missouri type (SEMD-MO) [MIM:602111]. A bone disease characterized by moderate to severe metaphyseal changes, mild epiphyseal involvement, rhizomelic shortening of the lower limbs with bowing of the femora and/or tibiae, coxa vara, genu varum and pear-shaped vertebrae in childhood. Epimetaphyseal changes improve with age.
    Defects in MMP13 are the cause of metaphyseal anadysplasia type 1 (MANDP1) [MIM:602111]. Metaphyseal anadysplasia consists of an abnormal bone development characterized by severe skeletal changes that, in contrast with the progressive course of most other skeletal dysplasias, resolve spontaneously with age. Clinical characteristics are evident from the first months of life and include slight shortness of stature and a mild varus deformity of the legs. Patients attain a normal stature in adolescence and show improvement or complete resolution of varus deformity of the legs and rhizomelic micromelia.
  • 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

    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Alternative names

    • Collagenase 3
    • Matrix metalloproteinase-13
    • MMP-13
    • Mmp13
    • MMP13_HUMAN
    see all

Images

  • Slope=V=7.00E-03 OD/min

  • control slope = 7.00E-03 OD/min

    inhibitor (100nM) slope = 4.50E-04 OD/min

    inhibitor % activity remaining = (4.50E-04 / 7.00E-03) x 100 = 6.4%

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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