Scientific Publications

Refined Preparation and Use of Anti-K- epsilon -GG Antibody Enables Routine Quantification of 10,000s of Ubiquitination Sites in Single Proteomics Experiments.

Publication TypeJournal Article
AuthorsUdeshi, ND, Svinkina T., Mertins P., Kuhn E., Mani D. R., Qiao JW, and Carr SA
AbstractDetection 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.
Year of Publication2012
JournalMolecular & cellular proteomics : MCP
Date Published (YYYY/MM/DD)2012/12/24
ISSN Number1535-9476
DOI10.1074/mcp.O112.027094
PubMedhttp://www.ncbi.nlm.nih.gov/pubmed/23266961?dopt=Abstract