title: Detection of Connexins in Liver Cells Using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis and Immunoblot Analysis.
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.
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarkbeeklaan 103, 1090, Jette, Brussel, Belgium. jwillebr@vub.ac.be.
authors.forename: Joost
authors.initials: J
authors.lastname: Willebrords
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarkbeeklaan 103, 1090, Jette, Brussel, Belgium.
authors.forename: Michaël
authors.initials: M
authors.lastname: Maes
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarkbeeklaan 103, 1090, Jette, Brussel, Belgium.
authors.forename: Sara Crespo
authors.initials: SC
authors.lastname: Yanguas
authors.affiliation: Department of Pathology, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP, Brazil.
authors.forename: Bruno
authors.initials: B
authors.lastname: Cogliati
authors.affiliation: Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarkbeeklaan 103, 1090, Jette, Brussel, Belgium.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Vinken
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D015153:Blotting, Western / Q000379:methods*; D003101:Collodion; D017630:Connexins / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D022781:Hepatocytes / Q000378:metabolism*; D051379:Mice; D051381:Rats; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
chemical_list: D000906:Antibodies; D017630:Connexins; D012967:Sodium Dodecyl Sulfate; D003101:Collodion
keywords: Antibody; Connexins; Enhanced chemiluminescence; Immunoblot analysis; Protein extraction; Sodium dodecyl sulfate polyacrylamide gel electrophoresis