Humana Press eBooks · 2006 · 14 citations · 13 references
Life SciencesBiochemistryMolecular BiologyMass Spectrometric AnalysisProtein BiochemistryProteomicsBottom-up ProteomicsMass Spectrometry-based ProteomicsProtein SeparationGel ElectrophoresisMatrix-assisted Laser Desorption/ionizationPolyacrylamide Gel ElectrophoresisProtein ExtractionProtein Sample PreparationSodium Dodecyl Sulfate-polyacrylamide Gel ElectrophoresisTwo-dimensional ElectrophoresisTwo-dimensional Gel Electrophoresis
Proteomics is the study of the set of proteins, or proteome, expressed by a cell under specific conditions. Proteomics methodology consists of protein extraction, protein separation, and protein identification. Currently, two-dimensional gel electrophoresis (2DE) and matrix-assisted laser-desorption ionization time of flight mass spectrometry are the most widespread methods for proteomic studies. The recent introduction of precast immobilized pH gradient gel strips, precast gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, and well-designed electrophoresis equipment has made 2DE a highly reproducible and relatively simple method for protein separation. Inherent limitations of the procedure, however, require approaches in sample preparation that may be cell- or tissue-dependent. This chapter describes a methodology for proteomic analysis of human neutrophils and discusses its applications.
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Quantitative analysis of complex protein mixtures using isotope-coded affinity tags
Steven P. Gygi, Beate Rist, Scott A. Gerber et al. · Nature Biotechnology · 1999 · 4.7K citations