Locating sites of Phosphotyrosine

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.