title: A combined free flow electrophoresis and DIGE approach to compare proteins in complex biological samples.
abstract: Free flow electrophoresis (FFE) has been applied in numerous studies as a protein separation technique due to its multiple advantages such as fast and efficient sample recovery, high resolving power, high reproducibility, and wide applicability to protein classes. As a stand-alone platform however, its utility in comparative proteomic analysis is limited as protein samples must be run sequentially rather than simultaneously which introduces inherent variability when attempting to perform quantitative analysis. Here we describe an approach combining fluorescent CyDye technology (DIGE) with FFE to simultaneously separate and identify differentially expressed proteins in a model cell system.
authors.affiliation: CSIRO, Preventative Health Flagship, Australia, Adelaide BC, SA, Australia. kim.fung@csiro.au
authors.forename: Kim Y C
authors.initials: KY
authors.lastname: Fung
authors.forename: Chris
authors.initials: C
authors.lastname: Cursaro
authors.forename: Tanya
authors.initials: T
authors.lastname: Lewanowitsch
authors.forename: Leah
authors.initials: L
authors.lastname: Cosgrove
authors.forename: Peter
authors.initials: P
authors.lastname: Hoffmann
mesh_terms: D002021:Buffers; D002232:Carbocyanines / Q000737:chemistry; D002457:Cell Extracts / Q000737:chemistry / Q000302:isolation & purification*; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D003720:Densitometry; D005456:Fluorescent Dyes / Q000737:chemistry; D019073:HT29 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D007526:Isoelectric Point; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D013194:Staining and Labeling
chemical_list: D002021:Buffers; D002232:Carbocyanines; D002457:Cell Extracts; D005456:Fluorescent Dyes; D011506:Proteins; C089187:cyanine dye 3; C085321:cyanine dye 5; C011203:cyanine dye 2