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


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.
Number of pages12
ISBN (Print)9781493911417
StatePublished - 2014
Externally publishedYes

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Antibody
  • Mass spectrometry
  • Quantitative proteomics
  • Ubiquitin

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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