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A combined free‐flow electrophoresis and DIGE approach to identify proteins regulated by butyrate in HT29 cells
Author(s) -
Fung Kim Y. C.,
Cursaro Chris,
Lewanowitsch Tanya,
Brierley Gemma V.,
McColl Shaun R.,
Lockett Trevor,
Head Richard,
Hoffmann Peter,
Cosgrove Leah
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000429
Subject(s) - proteome , free flow electrophoresis , difference gel electrophoresis , proteomics , western blot , butyrate , chemistry , biology , gel electrophoresis , microbiology and biotechnology , biochemistry , polyacrylamide gel electrophoresis , gel electrophoresis of proteins , enzyme , fermentation , gene
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.