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A combined free-flow electrophoresis and DIGE approach to identify proteins regulated by butyrate in HT29 cells.

Abstract
Many biologically active agents exert a pleiotropic response in cells and tissues. This presents challenges in descriptive and comparative analysis of the proteome in response to these agents. Although free-flow electrophoresis has been applied in a number of proteomic studies as a protein separation technique, the combination of free-flow electrophoresis and DIGE has not yet been investigated for comparative proteomic analysis. In this study, we have compared the effects of butyrate on HT29 colorectal cancer cells with a particular focus on apoptosis and describe the utility of a novel approach combining free-flow electrophoresis with DIGE to identify differentially expressed proteins. We verify the results obtained by the combined free-flow electrophoresis and DIGE approach with Western blot analysis of selected proteins. We also report for the first time the regulation of a number of proteins by butyrate in HT29 colorectal cells including peptidyl-prolyl cis-trans isomerase A (cyclophilin A) and profilin-1.
AuthorsKim Y C Fung, Chris Cursaro, Tanya Lewanowitsch, Gemma V Brierley, Shaun R McColl, Trevor Lockett, Richard Head, Peter Hoffmann, Leah Cosgrove
JournalProteomics (Proteomics) Vol. 11 Issue 5 Pg. 964-71 (Mar 2011) ISSN: 1615-9861 [Electronic] Germany
PMID21280223 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Chemical References
  • Butyrates
  • Profilins
  • Protective Agents
  • Proteome
  • Cyclophilin A
Topics
  • Apoptosis (drug effects)
  • Blotting, Western
  • Butyrates (pharmacology)
  • Cell Line, Tumor
  • Colorectal Neoplasms (genetics, metabolism, prevention & control)
  • Cyclophilin A (genetics, metabolism)
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic (drug effects)
  • HT29 Cells
  • Humans
  • Profilins (genetics, metabolism)
  • Protective Agents (pharmacology)
  • Proteome (genetics, metabolism)
  • Proteomics (methods)
  • Two-Dimensional Difference Gel Electrophoresis

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