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技术文章首页 > 技术文章 > 高压辅助in-gel蛋白酶解:定性和定量方面
高压辅助in-gel蛋白酶解:定性和定量方面
点击次数:856 发布时间:2009-07-06

    胰蛋白酶解仍然是一个蛋白质组学的基本工作流程。zui近几年,一些提高蛋白酶解的方法已经开发起来。尽管如此,仍然存在着新蛋白酶解方法的开发空间,以满足蛋白质组学中定性和定量的需求。另一个蛋白质组学的组成部分是蛋白质的分离作用。所有蛋白质组学中的分离技术大致可分为三组:凝胶基(gel-based),液相基(liquid phase-based)和混合型液相凝胶基(hybrid liquid phase-gel-based)。zui近,我们制订了一个综合方法: 利用直接目标胰蛋白酶解结合反相液相色谱法(RP-HPLC)分 离蛋白质( 2 ) 。在这里,我们报告利用压力循环技术( PCT )结合in-gel胰蛋白酶解和评估利用1DE来这个方法应用中以1DE分离的标记蛋白质进行定量。 

 

High pressure-assisted in-gel tryptic digestion: qualitative and quantitative aspects

Tryptic digestion of proteins continues to be a fundamental part of any proteomics workflow. A number of enhanced digestion protocols have been developed in recent years (1,2). Nonetheless, a need still exists for new digestion approaches that meet the demands of qualitative and quantitative proteomics. Another integral part of proteomics is separation of proteins. All separation techniques in proteomics may be broadly classified under three groups: gel-based, liquid phase-based and hybrid liquid phase-gel- based. Recently we have developed an integrated approach combining RP-HPLC separation of proteins with direct on target tryptic digestion (2). Here we report an application of pressure cycling technology (PCT) with tryptic in-gel- digestion of proteins and an evaluation of the application of this method to label-free quantitation of proteins separated by 1DE.


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