Analysis of the Global Changes in SH2 Binding Properties Using Mass Spectrometry Supported by Quantitative Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Technique

Methods Mol Biol. 2017:1555:419-428. doi: 10.1007/978-1-4939-6762-9_24.

Abstract

Quantitative mass spectrometry (MS)-based proteomics enables fast and reliable analysis of protein complexes. Its robustness and sensitivity effectively substitute traditional antibody-based approaches. Here, we describe the combination of mass spectrometry and Stable Isotope Labeling by Amino acids in Cell culture (SILAC) in characterization of the SH2 domain binding capacity.

Keywords: Peptide pull down; Quantitative mass spectrometry; SH2 domains; SILAC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry*
  • Cell Culture Techniques*
  • HeLa Cells
  • Humans
  • Isotope Labeling* / methods
  • Mass Spectrometry* / methods
  • Peptides / chemical synthesis
  • Peptides / chemistry
  • Protein Binding
  • Proteins / chemistry*
  • Proteins / metabolism
  • Proteomics / methods
  • Recombinant Fusion Proteins
  • Workflow
  • src Homology Domains*

Substances

  • Amino Acids
  • Peptides
  • Proteins
  • Recombinant Fusion Proteins