An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry

Dragana Lagundžin; Kimiko L. Krieger; Henry C.H. Law; Nicholas T. Woods

doi:10.1016/j.xpro.2022.101234

An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry

Dragana Lagundžin, Kimiko L. Krieger, Henry C.H. Law, Nicholas T. Woods

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

This protocol represents an optimized proteomics-based protocol for the endogenous protein enrichment and protein-protein interaction analysis. This 2-step protocol consists of: 1) co-immunoprecipitation of the bait protein; 2) the bait-protein interactions analysis using LC-MS/MS. Here, we used Dynabeads® for the enrichment of the target protein (the bait) and its interactors. We have tested the protocol using several different cell lines. Our conclusion is that the protocol is applicable to different cell lines and species. For complete details on the use and execution of this protocol, please refer to Lagundžin et al. (2019).

Original language	English (US)
Article number	101234
Journal	STAR Protocols
Volume	3
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2022.101234
State	Published - Mar 18 2022

Keywords

Mass Spectrometry
Protein Biochemistry
Protein expression and purification
Proteomics

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Neuroscience
General Immunology and Microbiology
General Medicine

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10.1016/j.xpro.2022.101234

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abstract = "This protocol represents an optimized proteomics-based protocol for the endogenous protein enrichment and protein-protein interaction analysis. This 2-step protocol consists of: 1) co-immunoprecipitation of the bait protein; 2) the bait-protein interactions analysis using LC-MS/MS. Here, we used Dynabeads{\textregistered} for the enrichment of the target protein (the bait) and its interactors. We have tested the protocol using several different cell lines. Our conclusion is that the protocol is applicable to different cell lines and species. For complete details on the use and execution of this protocol, please refer to Lagund{\v z}in et al. (2019).",

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AU - Lagundžin, Dragana

AU - Krieger, Kimiko L.

AU - Law, Henry C.H.

AU - Woods, Nicholas T.

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AB - This protocol represents an optimized proteomics-based protocol for the endogenous protein enrichment and protein-protein interaction analysis. This 2-step protocol consists of: 1) co-immunoprecipitation of the bait protein; 2) the bait-protein interactions analysis using LC-MS/MS. Here, we used Dynabeads® for the enrichment of the target protein (the bait) and its interactors. We have tested the protocol using several different cell lines. Our conclusion is that the protocol is applicable to different cell lines and species. For complete details on the use and execution of this protocol, please refer to Lagundžin et al. (2019).

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KW - Protein expression and purification

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An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry

Abstract

Keywords

ASJC Scopus subject areas

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