Non-covalent and covalent protein labeling in two-dimensional gel electrophoresis.
Over the past decades, several sensitive post-electrophoretic stains have been developed for an identification of proteins in general, or for a specific detection of post-translational modifications such as phosphorylation, glycosylation or oxidation. Yet, for a visualization and quantification of protein differences, the differential two-dimensional gel electrophoresis, termed DIGE, has become the method of choice for a detection of differences in two sets of proteomes. The goal of this review is to evaluate the use of the most common non-covalent and covalent staining techniques in 2D electrophoresis gels, in order to obtain maximal information per electrophoresis gel and for an identification of potential biomarkers. We will also discuss the use of detergents during covalent labeling, the identification of oxidative modifications and review influence of detergents on finger prints analysis and MS/MS identification in relation to 2D electrophoresis.
Riederer
Beat M
BM
Center for Psychiatric Neuroscience, Proteomic Unit, University Hospital of Lausanne (CHUV), 1008 Prilly-Lausanne, Switzerland. Beat.Riederer@chuv.ch
D000595:Amino Acid Sequence; D000818:Animals; D003902:Detergents; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013194:Staining and Labeling
D003902:Detergents; D005456:Fluorescent Dyes; D011506:Proteins