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Zymography Principles

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Zymography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1626))

Abstract

Zymography, the detection, identification, and even quantification of enzyme activity fractionated by gel electrophoresis, has received increasing attention in the last years, as revealed by the number of articles published. A number of enzymes are routinely detected by zymography, especially with clinical interest. This introductory chapter reviews the major principles behind zymography. New advances of this method are basically focused towards two-dimensional zymography and transfer zymography as will be explained in the rest of the chapters. Some general considerations when performing the experiments are outlined as well as the major troubleshooting and safety issues necessary for correct development of the electrophoresis.

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References

  1. Lantz MS, Ciborowski P (1994) Zymographic techniques for detection and characterization of microbial proteases. Methods Enzymol 235:563–594

    Article  CAS  PubMed  Google Scholar 

  2. Manchenko GP (2003) Handbook of detection of enzymes on electrophoretic gels, 2nd edn. CRC Press, USA, pp 1–39

    Google Scholar 

  3. Vandooren J, Geurts N, Martens E, Van den Steen PE, Opdenakker G (2013) Zymography methods for visualizing hydrolytic enzymes. Nat Methods 10:211–220

    Article  CAS  PubMed  Google Scholar 

  4. Wilkesman J, Kurz L (2009) Protease analysis by zymography: a review on techniques and patents. Recent Pat Biotechnol 3:175–184

    Article  CAS  PubMed  Google Scholar 

  5. Keow JY, Herrmann KM, Crawford BD (2011) Differential in vivo zymography: a method for observing matrix metalloproteinase activity in the zebrafish embryo. Matrix Biol 30:169–177

    Article  CAS  PubMed  Google Scholar 

  6. Sanaullah M, Razavi BS, Blagodatskaya E et al (2016) Biol Fertil Soils 52:505. doi:10.1007/s00374-016-1094-8

    Article  CAS  Google Scholar 

  7. Spohn M, Carminati A, Kuzyakov Y (2013) Soil zymography A novel in situ method for mapping distribution of enzyme activity in soil. Soil Biol Biochem 58:275–280

    Article  CAS  Google Scholar 

  8. Pan D, Hill AP, Kashou A, Wilson KA, Tan-Wilson A (2011) Electrophoretic transfer protein zymography. Anal Biochem 411:277–283

    Article  CAS  PubMed  Google Scholar 

  9. Wilkesman JG, Schröder HC (2007) Analysis of serine proteases from marine sponges by 2-D zymography. Electrophoresis 28:429–436

    Article  CAS  PubMed  Google Scholar 

  10. Snoek-van Beurden PA, Von den Hoff JW (2005) Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors. Biotechniques 38:73–83

    Article  CAS  PubMed  Google Scholar 

  11. Awde S, Marty-Gasset N, Wilkesman J, Rémignon H (2013) Proteolytic activity alterations resulting from force-feeding in Muscovy and Pekin ducks. Poult Sci 92:2997–3002

    Article  CAS  PubMed  Google Scholar 

  12. Rossano R, Larocca M, Riccio P (2011) 2-D zymographic analysis of Broccoli (Brassica oleracea L. var. Italica) florets proteases: follow up of cysteine protease isotypes in the course of post-harvest senescence. J Plant Physiol 168:1517–1525

    Article  CAS  PubMed  Google Scholar 

  13. Rothe GM (1994) Electrophoresis of enzymes, laboratory methods. Springer, New York

    Book  Google Scholar 

  14. Yasumitsu H, Ozeki Y, Kanaly RA (2016) RAMA casein zymography: time-saving and highly sensitive casein zymography for MMP7 and trypsin. Electrophoresis 37:2959–2962

    Article  CAS  PubMed  Google Scholar 

  15. Kupai K, Szucs G, Cseh S, Hajdu I, Csonka C, Csont T, Ferdinandy P (2010) Matrix metalloproteinase activity assays: importance of zymography. J Pharmacol Toxicol Methods 61:205–209

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We would like to thank Prof. Dr. J. Bubis and M. Calabokis (University Simón Bolívar) for their unconditional support during all this time. We thank Prof. Dr. W.E.G. Müller and H.C. Schröder, for opening the doors of their labs to host our research stay. We appreciate the support of the University of Carabobo.

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Correspondence to Jeff Wilkesman .

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Wilkesman, J., Kurz, L. (2017). Zymography Principles. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_1

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  • DOI: https://doi.org/10.1007/978-1-4939-7111-4_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7109-1

  • Online ISBN: 978-1-4939-7111-4

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