Post Translational Modification (PTMs)
Identify and localize PTMs such as phosphorylation, acetylation and oxidation with and without enrichment strategies.
Building on protein identification presented above (1), PTM analysis for modifications of peptides including Phosphorylation, Oxidation, Acetylation, Methylation (mono, di, tri), Ubiquitination, Biotinylation, Glutathionylation and a variety of other moieties is offered. Depending on a range of factors, enrichment(s) may or may not be necessary. In the case of phosphorylation, if you are studying a relatively pure protein in vitro in many instances we will be able to identify, localize and quantify (semi-quantitative) the occupancy rate without enrichments. In the case of global phospho-analysis we rely on enrichment technologies based on TiO2 as presented in Bodenmiller et. al Nat. Meth. (2007) 4: 231-7. Because of the paucity of phosphorylation and other modifications, large amounts of starting material are typically needed for global analysis (>2mg total protein). Other enrichment kits we have experience with and support come from various vendors including Cell Signaling Technology (CST), Promega, Thermo Fisher, Millipore, Protea and Sigma Aldrich. Please contact Ryan Leib for more information regarding PTM analysis, necessary protocols and/or kits.