title: Trends in sample preparation for classical and second generation proteomics.
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.
authors.affiliation: Dept. Química Analítica, Facultad de CC, Químicas, UCM, Av.Complutense s/n, Madrid 28040, Spain. bcanasmo@quim.ucm.es
authors.forename: Benito
authors.initials: B
authors.lastname: Cañas
authors.forename: Carmen
authors.initials: C
authors.lastname: Piñeiro
authors.forename: Enrique
authors.initials: E
authors.lastname: Calvo
authors.forename: Daniel
authors.initials: D
authors.lastname: López-Ferrer
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Gallardo
mesh_terms: D053000:Analytic Sample Preparation Methods / Q000639:trends*; D000818:Animals; D005591:Chemical Fractionation; D002853:Chromatography, Liquid / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D009862:Online Systems; D010447:Peptide Hydrolases / Q000378:metabolism; D011492:Protein Hydrolysates / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D011492:Protein Hydrolysates; D011506:Proteins; D011933:Reagent Kits, Diagnostic; D010447:Peptide Hydrolases