Detection of protein stoichiometric phosphorylation using Phos-tag sds-page

Abstract

Protein phosphorylation plays an important role in many cellular signaling which are relating to many diseases. Therefore, a variety of biochemical techniques has been developed to study protein phosphorylation in cells. Protein phosphorylation has traditionally been detected by radioisotope phosphate labeling of proteins with radioactive ATP. Phosphorylation sitc-specific antibodies are now available for the analysis of phosphorylation status at target sites. However, these antibodies cannot be used to detect unidentified phosphorylation sites. Recently, the Phos-tag technology has been developed to overcome the disadvantages and limitations of these methods. Phos-tag and its derivatives conjugated to biotin, acrylamide, or agarose, and can capture phosphate mono ester dianions bound to serine. threonine, and tyrosine residues, in an amino acid sequence-independent manner.