A simple and effective method for detecting phosphopeptides for phosphoproteomic analysis.

An immuno-dot blot method for detecting phosphopeptides is described. This method allows detection of phosphopeptides prior to analysis by mass spectrometry, and does not require specialized equipment. Further, this method allows peptides containing a given phosphoamino acid to be selectively detected by using phosphoamino acid-specific antibody. This phosphopeptide assay should be useful for researchers who do not have direct access to the mass spectrometers to prepare high quality samples by optimizing phosphopeptide enrichment procedures prior to mass spectrometry analysis.