Use of antibodies for detection of phosphorylated proteins separated by two-dimensional gel electrophoresis

Abstract

Protein phosphorylation and dephosphorylation are key regulatory mechanisms in prokaryotic and eukaryotic cells. Considering the role of phosphorylation in many human diseases, it appears a major challenge to refine on the methods to analyze the phospho-proteome. Here we review the use of monoclonal antibodies directed against specific phosphorylated amino acid residues to visualize phosphoproteins separated by two-dimensional gel electrophoresis. Strategies are described how this method can successfully be applied to create phospho-proteome maps of mammalian cells.