Trends in sample preparation for classical and second generation proteomics.
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.
Cañas
Benito
B
Dept. Química Analítica, Facultad de CC, Químicas, UCM, Av.Complutense s/n, Madrid 28040, Spain. bcanasmo@quim.ucm.es
Piñeiro
Carmen
C
Calvo
Enrique
E
López-Ferrer
Daniel
D
Gallardo
Jose Manuel
JM
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
D011492:Protein Hydrolysates; D011506:Proteins; D011933:Reagent Kits, Diagnostic; D010447:Peptide Hydrolases