Towards multidimensional liquid chromatography separation of proteins using fluorescence and isotope-coded protein labelling for quantitative proteomics.
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.
Tribl
Florian
F
Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
Lohaus
Christiane
C
Dombert
Tanja
T
Langenfeld
Elmar
E
Piechura
Heike
H
Warscheid
Bettina
B
Meyer
Helmut E
HE
Marcus
Katrin
K
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
D005456:Fluorescent Dyes; D007554:Isotopes; D020543:Proteome