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基于凝胶和非凝胶的蛋白质组学技术。

Gel-based and gel-free proteomic technologies.

作者信息

Scherp Peter, Ku Ginger, Coleman Liana, Kheterpal Indu

机构信息

Proteomics and Metabolomics Core, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA.

出版信息

Methods Mol Biol. 2011;702:163-90. doi: 10.1007/978-1-61737-960-4_13.

Abstract

Proteomics refers to the analysis of expression, localization, functions, posttranslational modifications, and interactions of proteins expressed by a genome at a specific condition and at a specific time. Mass spectrometry (MS)-based proteomic methods have emerged as a key technology for unbiased systematic and high-throughput identification and quantification of complex protein mixtures. These methods have the potential to reveal unknown and novel changes in protein interactions and assemblies that regulate cellular and physiological processes. Both gel-based (one-dimensional [1D] gel electrophoresis, two-dimensional [2D] polyacrylamide gel electrophoresis, 2D difference in-gel electrophoresis [DIGE]) and gel-free (liquid chromatography [LC], capillary electrophoresis) approaches have been developed and utilized in a variety of combinations to separate proteins prior to mass spectrometric analysis. Detailed protocols for global proteomic analysis from adipose-derived stem cells (ASCs) using two central strategies, 2D-DIGE-MS and 2D-LC-MS, are presented here.

摘要

蛋白质组学是指对基因组在特定条件和特定时间表达的蛋白质的表达、定位、功能、翻译后修饰及相互作用进行分析。基于质谱(MS)的蛋白质组学方法已成为无偏倚地系统、高通量鉴定和定量复杂蛋白质混合物的关键技术。这些方法有可能揭示调节细胞和生理过程的蛋白质相互作用和组装中未知的新变化。基于凝胶的方法(一维[1D]凝胶电泳、二维[2D]聚丙烯酰胺凝胶电泳、二维差异凝胶电泳[DIGE])和非凝胶方法(液相色谱[LC]、毛细管电泳)均已得到开发,并以各种组合方式用于在质谱分析之前分离蛋白质。本文介绍了使用两种核心策略,即二维差异凝胶电泳-质谱(2D-DIGE-MS)和二维液相色谱-质谱(2D-LC-MS),对脂肪来源干细胞(ASC)进行整体蛋白质组分析的详细方案。

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