Locating sites of Phosphotyrosine
The right hand Figure shows two work flows utilizing anti-phosphotyrosine antibodies to locate sites of tyrosine. The left hand workflow enriches for intact proteins containing phosphotyrosine, followed by 1D SDS PAGE, digestion of gel bands, TiO2 enrichment, mass spectrometric analysis and database searching.
The far right hand work flow enriches for phosphotyrosine peptides after digestion and prior to a Zip Tip clean up step, mass spectrometric analysis and database searching. The antibody pull down step is typically done in the investigator’s lab with sample submission to the MS & Proteomics Resource at the digest, Zip Tip, TiO2
, or mass spectrometric steps.