Some improvements in two-dimensional gel electrophoresis of proteins. Protein mapping of eukaryotic tissue extracts

David L. Wilson, Michael E. Hall, George C. Stone, Robert W. Rubin

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Abstract

In the course of adapting O'Farrell's (1975, J. Biol. Chem.250, 4007-4021) two-dimensional separation technique for proteins to eukaryotic material, we have made some modifications. During sample preparation, sodium dodecyl sulfate (SDS) can be included, with a resulting enhancement in reproducibility of gel patterns. However, heating in the presence of SDS leads to artifactual spots in the gels, probably as a result of protein charge modifications. Ultracentrifugation reduces the clogging at the top of the isoelectric focussing gel. For electrophoresis, some modifications of apparatus and technique are suggested. For the analysis of gels, a simple high-efficiency method for the counting of radioactivity in spots from dried gel slabs is described. In addition, an inexpensive microdensitometer option is described for the analysis of the autoradiographs. Patterns of proteins obtained from superior cervical sympathetic ganglia of rats and from other eukaryotic tissues are illustrated. Finally, a few of the proteins commonly found in mammalian tissue are identified on the gels.

Original languageEnglish (US)
Pages (from-to)33-44
Number of pages12
JournalAnalytical Biochemistry
Volume83
Issue number1
DOIs
StatePublished - Nov 1977

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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