ITRAQ-based shotgun neuroproteomics

Tong Liu, Jun Hu, Hong Li

Research output: Chapter in Book/Report/Conference proceedingChapter

22 Scopus citations

Abstract

Shotgun proteomics involves the analysis of peptides obtained by enzymatic digestions of proteins and subsequent identification via tandem mass spectrometry. This approach is an effective method for studying global protein expression in neuronal systems. The method described here is a quantitative shotgun neuroproteomics method using amine-specific isobaric tags for a relative and absolute quantitation (iTRAQ)-based workflow. We will provide the technical details for sample preparation, two-dimensional liquid chromatography, tandem mass spectrometry, database search, and statistical analysis to identify differentially expressed proteins. We will use a recent study on a rat model of multiple sclerosis, experimental autoimmune encephalomyelitis to illustrate the successful application of this method.

Original languageEnglish (US)
Title of host publicationNeuroproteomics
Subtitle of host publicationMethods and Protocols
EditorsAndrew Ottens, Kevin Wang, Kevin Wang
Pages201-216
Number of pages16
DOIs
StatePublished - 2009

Publication series

NameMethods in Molecular Biology
Volume566
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • EAE
  • Liquid chromatography
  • Multiple sclerosis
  • Posttranslational modification
  • Shotgun proteomics
  • Tandem mass spectrometry
  • iTRAQ

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