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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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Fundamentals of Zymography: Applications to the Study of Biological Samples

  • 实验方案

  • 发表日期：

    2025年

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  • Jorge Recio-Aldavero,
  • Laura Muñoz-Moreno,
  • I. Dolores Román,
  • Ana M. Bajo

摘要

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.