title: Evaluation of SYPRO Ruby total protein stain for the normalization of two-dimensional Western blots.
abstract: Due to post-translational modifications such as phosphorylation, proteins exist as distinct charge variants. Two-dimensional (2D) gel electrophoresis followed by immunoblotting enables the detection of these isoforms. For their accurate relative quantitation in different samples, a loading control is necessary to compensate for technical errors such as imprecise sample loading or transfer. The study reveals that the combinatory approach of SYPRO Ruby and chemiluminescence-based 2D Western blot analysis exhibits high linearity and excellent reproducibility and is applicable for limited sample amounts.
authors: Institute of Animal Breeding and Genetics, Department of Biomedical Sciences, University of Veterinary Medicine, A-1210 Vienna, Austria; Institute of Biotechnology in Animal Production, Department of Agrobiotechnology (IFA Tulln), University of Natural Resources and Applied Life Sciences Vienna, A-3430 Tulln, Austria. Electronic address: birgit.steinberger@boku.ac.at.
authors: Birgit
authors: B
authors: Steinberger
authors: Institute of Animal Breeding and Genetics, Department of Biomedical Sciences, University of Veterinary Medicine, A-1210 Vienna, Austria.
authors: Gottfried
authors: G
authors: Brem
authors: Institute of Animal Breeding and Genetics, Department of Biomedical Sciences, University of Veterinary Medicine, A-1210 Vienna, Austria; Institute of Biotechnology in Animal Production, Department of Agrobiotechnology (IFA Tulln), University of Natural Resources and Applied Life Sciences Vienna, A-3430 Tulln, Austria.
authors: Corina
authors: C
authors: Mayrhofer
mesh_terms: D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D009942:Organometallic Compounds / Q000737:chemistry*
chemical_list: D009942:Organometallic Compounds; C515601:Sypro Ruby
keywords: Isoform quantitation; Normalization; SYPRO Ruby; Two-dimensional Western blot