Cited by 368Advanced Proteomics Techniques and ApplicationsBioinformatics and Genomic NetworksMass Spectrometry Techniques and Applications

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Affinity-purification coupled to mass spectrometry: basic principles and strategies

Dunham, Wade H., Mullin, Michael, Gingras, Anne‐Claude·2012·PROTEOMICS

DOI 10.1002/pmic.201100523

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Abstract

Identifying the interactions established by a protein of interest can be a critical step in understanding its function. This is especially true when an unknown protein of interest is demonstrated to physically interact with proteins of known function. While many techniques have been developed to characterize protein-protein interactions, one strategy that has gained considerable momentum over the past decade for identification and quantification of protein-protein interactions, is affinity-purification followed by mass spectrometry (AP-MS). Here, we briefly review the basic principles used in affinity-purification coupled to mass spectrometry, with an emphasis on tools (both biochemical and computational), which enable the discovery and reporting of high quality protein-protein interactions.

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Background
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Epitope Tagging
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Tandem Affinity Purification of Protein Complexes from Mammalian Cells by the Strep/FLAG (SF)-TAP Tag
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A modified tandem affinity purification strategy identifies cofactors of the Drosophila nuclear receptor dHNF4
Yang, P., Sampson, H. M., & Krause, H. M. (2006) · PROTEOMICS · doi:10.1002/pmic.200500230 · Cited by 44
Drug target deconvolution by chemical proteomics
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Recombineering, transfection, Western, IP and ChIP methods for protein tagging via gene targeting or BAC transgenesis
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Authors: Dunham, W. H., Mullin, M., & Gingras, A.
Affiliations: Mount Sinai Hospital (CA)
Corresponding author: Gingras, Anne‐Claude
ORCID: Dunham, Wade H.
Mullin, Michael
Gingras, Anne‐Claude
Venue: PROTEOMICS
Publisher: Wiley
Published: 2012-05
Volume: 12
Issue: 10
Pages: 1576-1590
ISSN: 1615-9853 (ISSN-L)
1615-9861
DOI: 10.1002/pmic.201100523
OpenAlex: W1991953772
Cited by: 368
References: 144
Citations by year: 2026: 16 · 2025: 33 · 2024: 29 · 2023: 24 · 2022: 27 · 2021: 33 · 2020: 24 · 2019: 31 · 2018: 27 · 2017: 37 · 2016: 19 · 2015: 23 · 2014: 27 · 2013: 13 · 2012: 4
FWCI: 1.99
Type: journal article
Language: en
Open access: closed
Licence: onlinelibrary.wiley.com/termsAndConditions#vor (vor, 49 d)
Locations: PROTEOMICS (publishedVersion)
PubMed (publishedVersion)
Funders: Canadian Institutes of Health Research (MOP 84314)
Topics: Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bioinformatics and Genomic Networks
Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
Keywords: affinity purification, protein-protein interaction, affinity purification mass spectrometry, protein-protein interaction network, protein complex identification
MeSH terms: Animals, Chromatography, Affinity, Humans, Proteins, Mass Spectrometry, Protein Interaction Mapping, Systems Biology

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Affinity-purification coupled to mass spectrometry: basic principles and strategies

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