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

MMP Activity Detection in Zymograms.

Author information

1
Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged, Hungary. bencsik.peter@med.u-szeged.hu.
2
Pharmahungary Group, Szeged, Hungary. bencsik.peter@med.u-szeged.hu.
3
Institute for Heart Research, Slovak Academy of Sciences, Bratislava, Slovak Republic.
4
Institute of Physiology, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic.
5
Department of Biochemistry, Faculty of Medicine, University of Szeged, Szeged, Hungary.
6
Pharmahungary Group, Szeged, Hungary.
7
Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary.

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

PMID:
28608200
DOI:
10.1007/978-1-4939-7111-4_6
[Indexed for MEDLINE]

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