title: Principles and examples of gel-based approaches for phosphoprotein analysis.
abstract: Methods for analyzing the phosphorylation status of proteins are essential to investigate in detail key cellular processes, including signal transduction and cell metabolism. The transience of this post-translational modification and the generally low abundance of phosphoproteins require specific enrichment and/or detection steps prior to analysis. Here, we describe three gel-based approaches for the analysis of differentially expressed phosphoproteins. These approaches comprise (1) the sequential fluorescence staining of two-dimensional (2-D) gels using Pro-Q(®) Diamond and SYPRO(®) Ruby dyes to visualize and quantify phosphoproteins in total cellular lysates as well as (2) affinity enrichment of phosphoproteins in conjunction with sequential fluorescence staining of the 2-D gels and (3) affinity enrichment of proteins prior to pre-electrophoretic fluorescence labeling and 2-D gel electrophoresis.
authors.affiliation: Institute of Animal Breeding and Genetics, University of Veterinary Medicine Vienna, Vienna, Austria.
authors.forename: Birgit
authors.initials: B
authors.lastname: Steinberger
authors.forename: Corina
authors.initials: C
authors.lastname: Mayrhofer
mesh_terms: D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D010750:Phosphoproteins / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods
chemical_list: D010750:Phosphoproteins; D020543:Proteome