In situ zymography.

Methods in Molecular Biology
Sarah J George, Jason L Johnson

Abstract

In situ zymography is a unique laboratory technique that enables the localisation of matrix-degrading metalloproteinase (MMP) activity in histological sections. Frozen sections are placed on glass slides coated with fluorescently labelled matrix proteins. After incubation MMP activity can be observed as black holes in the fluorescent background due to proteolysis of the matrix protein. Alternatively frozen sections can be incubated with matrix proteins conjugated to quenched fluorescein. Proteolysis of the substrate by MMPs leads to the release of fluorescence. This technique can be combined with immunohistochemistry to enable co-location of proteins such as cell type markers or other proteins of interest. Additionally, this technique can be adapted for use with cell cultures, permitting precise location of MMP activity within cells, time-lapse analysis of MMP activity and analysis of MMP activity in migrating cells.

Citations

Sep 13, 2013·Molecular Neurobiology·Mayank Chaturvedi, Leszek Kaczmarek
Sep 29, 2012·Journal of Neurochemistry·Lisa A RidnourCarol A Colton
Jul 24, 2012·In Vitro Cellular & Developmental Biology. Animal·Aastha ChhabraVibha Rani
Jul 23, 2013·Asian Pacific Journal of Tropical Medicine·Bao ShanZhuo-Ri Li
Apr 17, 2020·Frontiers in Immunology·Mordehay KlepfishIrit Sagi

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