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.
MeSH terms
-
Analytic Sample Preparation Methods / trends*
-
Animals
-
Chemical Fractionation
-
Chromatography, Liquid / methods
-
Electrophoresis, Gel, Two-Dimensional / methods
-
Gene Expression Profiling / methods
-
Humans
-
Online Systems
-
Peptide Hydrolases / metabolism
-
Protein Hydrolysates / isolation & purification
-
Proteins / isolation & purification*
-
Proteomics / methods*
-
Reagent Kits, Diagnostic
-
Spectrometry, Mass, Electrospray Ionization / methods
-
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
Substances
-
Protein Hydrolysates
-
Proteins
-
Reagent Kits, Diagnostic
-
Peptide Hydrolases