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Detection of Connexins in Liver Cells Using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis and Immunoblot Analysis

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Gap Junction Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1437))

Abstract

Since connexin expression is partly regulated at the protein level, immunoblot analysis represents a frequently addressed technique in the connexin research field. The present chapter describes the setup of an immunoblot procedure, including protein extraction and quantification from biological samples, gel electrophoresis, protein transfer, and immunoblotting, which is optimized for analysis of connexins in liver tissue. In essence, proteins are separated on a polyacrylamide gel using sodium dodecyl sulfate followed by transfer of proteins on a nitrocellulose membrane. The latter allows specific detection of connexins with antibodies combined with revelation through enhanced chemiluminescence.

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Acknowledgements

This work was financially supported by the University Hospital of the Vrije Universiteit Brussel-Belgium (Willy Gepts Fonds UZ-VUB), the Fund for Scientific Research-Flanders (FWO grants G009514N and G010214N), the European Research Council (ERC Starting Grant 335476), the University of São Paulo-Brazil, and the Foundation for Research Support of the State of São Paulo (FAPESP SPEC grant 2013/50420-6).

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Authors and Affiliations

  1. Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarkbeeklaan 103, 1090, Jette, Brussel, Belgium

    Joost Willebrords, Michaël Maes, Sara Crespo Yanguas & Mathieu Vinken

  2. Department of Pathology, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP, Brazil

    Bruno Cogliati

Authors
  1. Joost Willebrords
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  2. Michaël Maes
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  3. Sara Crespo Yanguas
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  4. Bruno Cogliati
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  5. Mathieu Vinken
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Corresponding author

Correspondence to Joost Willebrords .

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Editors and Affiliations

  1. Department of in Vitro Toxicology and D, Vrije Universiteit Brussel, Brussels, Belgium

    Mathieu Vinken

  2. BHF Glasgow Cardiovascular Research, University of Glasgow, Glasgow, United Kingdom

    Scott R. Johnstone

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Willebrords, J., Maes, M., Yanguas, S.C., Cogliati, B., Vinken, M. (2016). Detection of Connexins in Liver Cells Using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis and Immunoblot Analysis. In: Vinken, M., Johnstone, S. (eds) Gap Junction Protocols. Methods in Molecular Biology, vol 1437. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3664-9_3

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  • DOI: https://doi.org/10.1007/978-1-4939-3664-9_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3662-5

  • Online ISBN: 978-1-4939-3664-9

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