Evaluation of several two-dimensional gel electrophoresis techniques in cardiac proteomics

Zhao Bo Li, Paul Flint, Marvin O. Boluyt

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

Two-dimensional gel electrophoresis (2-DE) is currently the best method for separating complex mixtures of proteins, and its use is gradually becoming more common in cardiac proteome analysis. A number of variations in basic 2-DE have emerged, but their usefulness in analyzing cardiac tissue has not been evaluated. The purpose of the present study was to systematically evaluate the capabilities and limitations of several 2-DE techniques for separating proteins from rat heart tissue. Immobilized pH gradient strips of various pH ranges, parameters of protein loading and staining, subcellular fractionation, and detection of phosphorylated proteins were studied. The results provide guidance for proteome analysis of cardiac and other tissues in terms of selection of the isoelectric point separating window for cardiac proteins, accurate quantitation of cardiac protein abundance, stabilization of technical variation, reduction of sample complexity, enrichment of low-abundant proteins, and detection of phosphorylated proteins.

Original languageEnglish (US)
Pages (from-to)3572-3585
Number of pages14
JournalElectrophoresis
Volume26
Issue number18
DOIs
Publication statusPublished - Sep 2005
Externally publishedYes

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Keywords

  • Cardiac proteomics
  • Heart
  • Proteomics
  • Two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Clinical Biochemistry

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