MMP activity detection in zymograms

Bencsik, Péter and Bartekova, Monika and Görbe, Anikó and Kiss, Krisztina and Pálóczi, János and Radosinska, Jana and Szűcs, Gergő and Ferdinandy, Péter (2016) MMP activity detection in zymograms. In: Zymography. Methods in Molecular Biology . Springer-Verlag, pp. 53-70.

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Abstract

Matrix metalloproteinases (MMP) belong to a distinguished class of zinc-depending 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 co-polymerized 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 of several sample measurements.

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