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An Inexpensive Staining Alternative for Gelatin Zymography Gels

Center for Medical Research, Medical Faculty, Johannes Kepler University, 4020 Linz, Austria
Author to whom correspondence should be addressed.
Methods Protoc. 2019, 2(3), 61;
Received: 6 June 2019 / Revised: 8 July 2019 / Accepted: 23 July 2019 / Published: 24 July 2019
PDF [803 KB, uploaded 25 July 2019]


Zymography is a widely used electrophoretic method to determine proteolytic activities in samples from various sources. The method is based on copolymerizing a suitable protein substrate within a sodium dodecyl sulfate-polyacrylamide gel. Following electrophoretic separation of the protease containing samples and a suitable incubation period, degradation of the substrate can be visualized through staining with Coomassie blue. Sites of proteolysis become visible as white bands on a dark blue background. However, this staining protocol requires considerable amounts of ethanol and acetic acid to remove unbound dye molecules. In this report, we describe a new staining protocol using Ponceau S which offers substantial advantages in terms of assay usability and cost reduction, especially when performing large quantities of zymograms or in resource-limited settings. Fast and reproducible staining of zymograms with our protocol is demonstrated, and reliable quantitation of proteolytic activity in comparison to the standard Coomassie staining procedure is shown. View Full-Text
Keywords: gelatin zymography; Coomassie; Ponceau; inexpensive staining method gelatin zymography; Coomassie; Ponceau; inexpensive staining method

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Wechselberger, C.; Doppler, C.; Bernhard, D. An Inexpensive Staining Alternative for Gelatin Zymography Gels. Methods Protoc. 2019, 2, 61.

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