Phosphopeptides and phosphosites can be identified from proteins isolated in polyacrylamide gel slices. The approach is the same as used in the Protein –ID-gel service except that after digestion a titanium dioxide enrichment step is used to pull the phopshopeptides away from the non-phosphopeptides and thereby enrich this population of peptides. Both the enriched and un-enriched peptides are analyzed by LC-MS/MS separately.
Suitable Protein Complexity
|PTM-phospho-gel - <50 proteins||Less than 50 proteins||In gel digestion using a single enzyme, TiO2 phosphopeptide enrichment, LC-MS/MS analysis of both the enriched and non-enriched fractions (2 LC-MS/MS runs); short length gradient for <50 proteins and medium length gradient for <500 proteins, and a database search|
|PTM-phospho-gel - <500 proteins||Less than 500 proteins|
Kiraly, D.D., Stone, K.L., Colangelo, C.M., Abbott, T., Wang, Y., Mains, R.E., Eipper, B.A. (2011). Identification of Kalirin-7 as a Potential Post-Synaptic Density Signaling Hub. J Proteome Res. 10(6):2828-2841.
(PMCID: PMC3107868) (PMID: 21488700).
- Application Note 6: Using MD-Score and PhosphoRS in YPED to aid in localization and verification of phosphosites
- Sample Submission – YPED