Sample Preparation for Relative Quantitation of Proteins Using Tandem Mass Tags (TMT) and Mass Spectrometry (MS)

Methods Mol Biol. 2018:1741:135-149. doi: 10.1007/978-1-4939-7659-1_11.

Abstract

Quantitative proteome analysis allows comparisons of protein or phosphoprotein levels across multiple cell types or conditions. A number of experimental approaches have been described toward quantitative proteomics. In this chapter, we focus on Tandem Mass Tag (TMT) isobaric labeling of peptides for global, relative quantitation of proteins and phosphopeptides. To date, there has been no published protocol describing chemical labeling of small amounts of peptides specifically extracted from small tumor samples, for which rigorous sample preparation is necessary to ensure reproducible TMT labeling.

Keywords: MaxQuant; Peptide labeling; Phosphoproteomics; Proteomics; Relative quantitation of proteins; TMT; Tandem mass tags.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromatography, Liquid
  • Computational Biology / methods
  • Hydrogen-Ion Concentration
  • Hydrophobic and Hydrophilic Interactions
  • Peptides
  • Phosphopeptides / chemistry
  • Proteins / chemistry*
  • Proteome*
  • Proteomics* / methods
  • Software
  • Tandem Mass Spectrometry* / methods

Substances

  • Peptides
  • Phosphopeptides
  • Proteins
  • Proteome