Global ubiquitination analysis by SILAC in mammalian cells

Zhiping Wu, Chan Hyun Na, Haiyan Tan, Junmin Peng

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Ubiquitination is a versatile and dynamic posttranslational modification in cells, regulating almost all cellular events. With rapid developments of affinity capture reagents and high-resolution mass spectrometry, it is now feasible to globally analyze the ubiquitinated proteome (ubiquitome) using quantitative strategies, such as stable isotope labeling with amino acids in cell culture (SILAC). Here we describe in detail a SILAC protocol to profile the ubiquitome in mammalian cells including protein labeling, antibody-based enrichment, and analysis by mass spectrometry.

Original languageEnglish (US)
Title of host publicationStable Isotope Labeling by Amino Acids in Cell Culture (SILAC)
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages149-160
Number of pages12
ISBN (Print)9781493911417
DOIs
StatePublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1188
ISSN (Print)1064-3745

Keywords

  • Antibody
  • Mass spectrometry
  • Quantitative proteomics
  • SILAC
  • Ubiquitin

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Global ubiquitination analysis by SILAC in mammalian cells'. Together they form a unique fingerprint.

Cite this