Tribl Florian, Lohaus Christiane, Dombert Tanja, Langenfeld Elmar, Piechura Heike, Warscheid Bettina, Meyer Helmut E, Marcus Katrin
Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
Proteomics. 2008 Mar;8(6):1204-11. doi: 10.1002/pmic.200700736.
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.
高效液相色谱法（HPLC）已成为分离蛋白质的一种重要工具。为了应对复杂蛋白质组的分析及其内部蛋白质的定量变化，我们开发了一种基于多维液相色谱（MDLC）的方法，随后进行大型凝胶一维十二烷基硫酸钠聚丙烯酰胺凝胶电泳（1-D SDS-PAGE）。在此，我们提出了一种新颖的策略，该策略能够在经过三个分离和分级步骤后，同时鉴定和定量差异调节的蛋白质。这个MDLC平台整合了使用荧光和同位素编码标签进行双重蛋白质标记的优势，以便随后通过质谱法检测和定量蛋白质的丰度比。
