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

Date

2011

Authors

Fung, K.
Cursaro, V.
Lewanowitsch, T.
Brierley, G.
McColl, S.
Lockett, T.
Head, R.
Hoffmann, P.
Cosgrove, L.

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Journal article

Citation

Proteomics, 2011; 11(5):964-971

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Kim Y. C. Fung, Chris Cursaro, Tanya Lewanowitsch, Gemma V. Brierley, Shaun R. McColl, Trevor Lockett, Richard Head, Peter Hoffmann and Leah Cosgrove

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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.

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Article first published online: 31 JAN 2011

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Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

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