MMP Activity Detection in Zymograms

Methods Mol Biol. 2017;1626:53-70. doi: 10.1007/978-1-4939-7111-4_6.

Abstract

Matrix metalloproteinases (MMP) belong to a distinguished class of zinc-dependent endopeptidases. Zymography is a semi-quantitative tool for determining the activity of different MMP isoenzymes in a variety of biological samples. In substrate gel zymography, protein samples of different origin (tissue, cell lysates, plasma/serum, perfusates, other liquids) are separated in sodium dodecyl sulfate (SDS) polyacrylamide gels containing copolymerized substrate (gelatin, casein, elastin, etc.), and after incubation-enabling substrate cleavage by MMPs, MMP activities are detected after the gel staining as transparent bands against a dark-blue background. In situ zymography is a histological modification of substrate zymography in frozen sections, allowing detection of the localization of the MMP activities within the tissue. Here, we describe detailed experimental protocols of all abovementioned techniques and provide examples for several sample measurements.

Keywords: Casein; Gelatin; In situ zymography; Matrix metalloproteinase activity; Substrate zymography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caseins / metabolism
  • Cells, Cultured
  • Elastin / metabolism
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Enzyme Assays / methods*
  • Gelatin / metabolism
  • Humans
  • Matrix Metalloproteinases / analysis*
  • Matrix Metalloproteinases / metabolism
  • Staining and Labeling / methods
  • Substrate Specificity

Substances

  • Caseins
  • Gelatin
  • Elastin
  • Matrix Metalloproteinases