Trends in sample preparation for classical and second generation proteomics |
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Authors: | Cañas Benito Piñeiro Carmen Calvo Enrique López-Ferrer Daniel Gallardo Jose Manuel |
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Institution: | Dept. Química Analítica, Facultad de CC, Químicas, UCM, Av.Complutense s/n, Madrid 28040, Spain. bcanasmo@quim.ucm.es |
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Abstract: | 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. |
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