|Publication Type||Journal Article|
|Year of Publication||2012|
|Authors||Udeshi, ND, Svinkina, T, Mertins, P, Kuhn, E, Mani, DR, Qiao, JW, Carr, SA|
|Journal||Molecular & cellular proteomics : MCP|
Detection of endogenous ubiquitination sites by mass spectrometry has dramatically improved with the commercialization of anti-K-ε-GG remnant antibodies. Here we describe a number of improvements to the K-ε-GG enrichment workflow including optimized antibody and peptide input requirements, antibody crosslinking, and improved offline fractionation prior to enrichment. This refined and practical workflow enables routine identification and quantification of approximately 20,000 distinct endogenous ubiquitination sites in a single SILAC experiment using moderate amounts of protein input.