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Assessment of Gelatinases (MMP-2 and MMP-9) by Gelatin Zymography

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Metastasis Research Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 878))

Abstract

Gelatin zymography is a simple yet powerful method to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It is particularly useful for the assessment of two key members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. This polyacrylamide gel electrophoresis-based method can provide a reliable assessment of the type of gelatinase, relative amount, and activation status (latent, compared with active enzyme forms) in cultured cells, tissues, and biological fluids. The method can be used to investigate factors that regulate gelatinase expression and modulate zymogen activation in experimental systems. The system provides information on the pattern of gelatinase expression and activation in human cancer tissues and how this relates to cancer progression. Interpretation of the data obtained in gelatin zymography requires a thorough understanding of the principles and pitfalls of the technique; this is particularly important when evaluating enzyme levels and the presence of active gelatinase species. If properly used, gelatin zymography is an excellent tool for the study of gelatinases in biological systems.

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Acknowledgements

The authors would like to thank all the ex-members of the Fridman lab for suggestions and tips throughout the years. This work has been supported by an NIH/NCI grant (R01 CA-61986) to R.F.

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, Proteases and Cancer Program, Karmanos Cancer Institute, Wayne State University, Detroit, MI, USA

    Marta Toth & Anjum Sohail

  2. Department of Pathology, Wayne State University, School of Medicine, Detroit, MI, USA

    Rafael Fridman

Authors
  1. Marta Toth
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  2. Anjum Sohail
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  3. Rafael Fridman
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Corresponding author

Correspondence to Rafael Fridman .

Editor information

Editors and Affiliations

  1. School of Life Sciences, Molecular and Applied Biosciences, University of Westminster, New Cavendish Street 115, London, W1W 6UW, United Kingdom

    Miriam Dwek

  2. School of Life Sciences, Oxford Brookes University, Gipsy Lane, Oxford, OX3 0BP, United Kingdom

    Susan A. Brooks

  3. Inst. Anatomie und Experimentelle, Morphologie, Universitätsklinikum Hamburg, Martinistr. 52, Hamburg, 20246, Germany

    Udo Schumacher

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Toth, M., Sohail, A., Fridman, R. (2012). Assessment of Gelatinases (MMP-2 and MMP-9) by Gelatin Zymography. In: Dwek, M., Brooks, S., Schumacher, U. (eds) Metastasis Research Protocols. Methods in Molecular Biology, vol 878. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-854-2_8

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  • DOI: https://doi.org/10.1007/978-1-61779-854-2_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-853-5

  • Online ISBN: 978-1-61779-854-2

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