Enrichment strategies for posttranslational modifications
In order to determine the baseline signaling in cancer cell lines, to study the dynamic response of cell signaling upon different stimuli and to elucidate a drug’s cellular mode of action, we utilize various techniques for enrichment of posttranslational modifications.
An HPLC based Fe-IMAC column method developed in our lab enables us to comprehensively and reproducibly purify phosphopeptides out of complex lysates. Furthermore, we apply antibody-based immunoprecipitation approaches to specifically enrich for phosphorylated tyrosines and ubiquitinated or acetylated lysines.
These strategies facilitate the quantitative monitoring of thousands of modified peptides across different (treatment) conditions.