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

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

Author information

1
Department of Cancer Biology and Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, Massachusetts.
2
Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.

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.

KEYWORDS:

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

PMID:
30706993
PMCID:
PMC6579647
[Available on 2020-06-01]
DOI:
10.1002/cpps.84

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