Gel-aided sample preparation (GASP)-A simplified method for gel-assisted proteomic sample generation from protein extracts and intact cells

被引:88
作者
Fischer, Roman [1 ]
Kessler, Benedikt M. [1 ]
机构
[1] Univ Oxford, Nuffield Dept Med, Target Discovery Inst, TDI Mass Spectrometry Lab, Oxford OX3 7FZ, England
基金
英国医学研究理事会;
关键词
Bottom-up proteomics; Comparative proteomics; Gel-assisted sample preparation; Sample preparation; Technology; DIGESTION; IDENTIFICATION; TISSUES;
D O I
10.1002/pmic.201400436
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We describe a gel-assisted proteomic sample preparation method for MS analysis. Solubilized protein extracts or intact cells are copolymerized with acrylamide, facilitating denaturation, reduction, quantitative cysteine alkylation, and matrix formation. Gel-aided sample preparation has been optimized to be highly flexible, scalable, and to allow reproducible sample generation from 50 cells to milligrams of protein extracts. This methodology is fast, sensitive, easy-to-use on a wide range of sample types, and accessible to nonspecialists.
引用
收藏
页码:1224 / 1229
页数:6
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