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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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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