Refined preparation and use of anti-diglycine remnant (K-ε-GG) antibody enables routine quantification of 10,000s of ubiquitination sites in single proteomics experiments.
Mol Cell Proteomics
; 12(3): 825-31, 2013 Mar.
Article
em En
| MEDLINE
| ID: mdl-23266961
ABSTRACT
Detection of endogenous ubiquitination sites by mass spectrometry has dramatically improved with the commercialization of anti-di-glycine remnant (K-ε-GG) antibodies. Here, we describe a number of improvements to the K-ε-GG enrichment workflow, including optimized antibody and peptide input requirements, antibody cross-linking, and improved off-line fractionation prior to enrichment. This refined and practical workflow enables routine identification and quantification of â¼20,000 distinct endogenous ubiquitination sites in a single SILAC experiment using moderate amounts of protein input.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteoma
/
Proteômica
/
Ubiquitinação
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article