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Updated: May 10, 2025

Detection of Protease Activity by Fluorescent Peptide Zymography
Published on: January 20, 2019
Fundamentals of Zymography: Applications to the Study of Biological Samples
Jorge Recio-Aldavero1, Laura Muñoz-Moreno1, I Dolores Román1
1Research group: Cancers of Epithelial Origin. Facultad de Medicina y Ciencias de la Salud, Departamento de Biología de Sistemas, Unidad de Bioquímica y Biología Molecular, Campus Científico-Tecnológico, Universidad de Alcalá, Alcalá de Henares, 28805, Spain.
Abstract:
Zymography is a very useful technique to determine the gelatinolytic activity of matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9). This work describes the preparation and use of different biological samples (culture media, cell lysates, tissue extracts, and extracellular vesicles), as well as the determination of gelatinolytic activity in these samples. Proteins are separated on sodium dodecyl sulfate (SDS) polyacrylamide gels in the presence of gelatin. Once the total proteins have been resolved, specific activation of MMP-2 and MMP-9 found in the gel and staining-destination are performed to visualize this enzymatic activity. With a fast and simple method, it is possible to determine the catalytic activity of gelatinases 2 and 9 from different types of biological samples.
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