title: Towards multidimensional liquid chromatography separation of proteins using fluorescence and isotope-coded protein labelling for quantitative proteomics.
abstract: HPLC has emerged as a valuable tool for separating proteins. To address the analysis of complex proteomes and quantitative changes of proteins therein, we developed a multidimensional LC (MDLC)-based approach followed by large gel 1-D SDS-PAGE. Here we present a novel strategy that allows for simultaneously identifying and quantifying differentially regulated proteins following three separation and fractionation steps. This MDLC platform integrates advantages of dual protein labelling using both fluorescence and isotope-coded tags for subsequent detection and quantification of abundance ratios of proteins by MS.
authors.affiliation: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Tribl
authors.forename: Christiane
authors.initials: C
authors.lastname: Lohaus
authors.forename: Tanja
authors.initials: T
authors.lastname: Dombert
authors.forename: Elmar
authors.initials: E
authors.lastname: Langenfeld
authors.forename: Heike
authors.initials: H
authors.lastname: Piechura
authors.forename: Bettina
authors.initials: B
authors.lastname: Warscheid
authors.forename: Helmut E
authors.initials: HE
authors.lastname: Meyer
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D007553:Isotope Labeling / Q000379:methods*; D007554:Isotopes / Q000737:chemistry; D013058:Mass Spectrometry; D020543:Proteome / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D005456:Fluorescent Dyes; D007554:Isotopes; D020543:Proteome