Use of Gel Zymography to Examine Matrix Metalloproteinase (gelatinase) Expression in Brain Tissue or in Primary Glial Cultures

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2011 Dec 7

PMID 22144310

Citations 52

Authors

Harald Frankowski

Yu-Huan Gu

Ji Hoe Heo

Richard Milner

Gregory J Del Zoppo

Affiliations

Soon will be listed here.

Abstract

Glia synthesize, package, and secrete several species of matrix proteases, including the gelatinases (pro-)MMP-2 and (pro-)MMP-9. In appropriate settings (e.g., experimental ischemia), these MMPs can be assayed from cerebral tissues or from astrocytes and microglia in culture by enzymatic substrate-dependent assays and by gelatin-based zymography. We describe the methodologies for the sensitive quantitative development of the inactive and active forms of both MMP-2 and MMP-9 from tissues and cells, by means of lysis of the collagen substrate in collagen-impregnated gel electropheresis by the zymogen and active gelatinases. These methodologies are a refinement of those used commonly, with instructions to increase sensitivity. Serious and often overlooked issues regarding sources of sample contamination and elements confounding the MMP band development and their interpretation are discussed.

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