Trends in sample preparation for classical and second generation proteomics

被引:123
作者
Canas, Benito [1 ]
Pineiro, Carmen
Calvo, Enrique
Lopez-Ferrer, Daniel
Manuel Gallardo, Jose
机构
[1] UCM, Fac CC Quim, Dept Quim Analit, Av Complutense SN, Madrid 28040, Spain
[2] CSIC, IIM, C Eduardo Cabello 6, Vigo 36208, Spain
[3] CNIC, Unidad Proteom, Madrid 28029, Spain
[4] CBM SO, Prot Chem & Proteom Lab, Madrid 28049, Spain
关键词
proteomics; sample preparation; 2-DE; ESI; MALDI; protein digestion;
D O I
10.1016/j.chroma.2007.01.045
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Sample preparation is a fundamental step in the proteomics workflow. However, it is not easy to find compiled information updating this subject. In this paper, the strategies and protocols for protein extraction and identification, following either classical or second generation proteomics methodologies, are reviewed. Procedures for: tissue disruption, cell lysis, sample pre-fractionation, protein separation by 2-DE, protein digestion, mass spectrometry analysis, multidimensional peptide separations and quantification of protein expression level are described. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:235 / 258
页数:24
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