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Serine Protease Zymography: Low-Cost, Rapid, and Highly Sensitive RAMA Casein Zymography

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Zymography

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

Abstract

To detect serine protease activity by zymography, casein and CBB stain have been used as a substrate and a detection procedure, respectively. Casein zymography has been using substrate concentration at 1 mg/mL and employing conventional CBB stain. Although ordinary casein zymography provides reproducible results, it has several disadvantages including time-consuming and relative low sensitivity. Improved casein zymography, RAMA casein zymography, is rapid and highly sensitive. RAMA casein zymography completes the detection process within 1 h after incubation and increases the sensitivity at least by tenfold. In addition to serine protease, the method also detects metalloprotease 7 (MMP7, Matrilysin) with high sensitivity.

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Acknowledgment

The author is grateful to Mariko Kawasaki-Yasumitsu for her excellent support and also to members of Kamimachi Hospital. This work was supported by a 2010 Incentive Grant from Yokohama Academic Foundation.

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Authors and Affiliations

  1. Expert Laboratory for Life Environments (ELLE), Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama, 236-0027, Japan

    Hidetaro Yasumitsu

  2. Division of Pharmacy, Kamimach Hospital, 1-7-34, Kochi, 780-0901, Japan

    Hidetaro Yasumitsu

Authors
  1. Hidetaro Yasumitsu
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Correspondence to Hidetaro Yasumitsu .

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Editors and Affiliations

  1. Centre for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, University of Carabobo, Valencia, Venezuela

    Jeff Wilkesman

  2. Centre for Environmental, Biology and Chemistry Research Facultad de Ciencias y Tecnología, University of Carabobo, Valencia, Venezuela

    Liliana Kurz

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Yasumitsu, H. (2017). Serine Protease Zymography: Low-Cost, Rapid, and Highly Sensitive RAMA Casein Zymography. 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_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7111-4_2

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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

  • eBook Packages: Springer Protocols

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