Tandem Affinity Purification and Mass Spectrometry (TAP-MS) for the Analysis of Protein Complexes

Curr Protoc Protein Sci. 2019 Jun;96(1):e84. doi: 10.1002/cpps.84. Epub 2019 Feb 1.

Abstract

Affinity purification followed by mass spectrometry has become the technique of choice to identify binding partners in biochemical complexes isolated from a physiologic cellular context. In this report we detail our protocol for tandem affinity purification (TAP) primarily based on the use of the FLAG and HA peptide epitopes, with a particular emphasis on factors affecting yield and specificity, as well as steps to implement an automated version of the TAP procedure. © 2019 by John Wiley & Sons, Inc.

Keywords: TAP; affinity purification; epitope tags; mass spectrometry; protein complex; protein-protein interaction.

Publication types

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

MeSH terms

  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Oligopeptides / chemistry
  • Proteomics
  • Retroviridae / chemistry
  • Retroviridae / genetics
  • Retroviridae / isolation & purification*
  • Tandem Affinity Purification / methods*
  • Tandem Mass Spectrometry / methods*

Substances

  • Oligopeptides
  • FLAG peptide