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

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 Dyn...

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Detalles Bibliográficos
Autores principales: Lagundžin, Dragana, Krieger, Kimiko L., Law, Henry C.-H., Woods, Nicholas T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8920916/
https://www.ncbi.nlm.nih.gov/pubmed/35300004
http://dx.doi.org/10.1016/j.xpro.2022.101234
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author Lagundžin, Dragana
Krieger, Kimiko L.
Law, Henry C.-H.
Woods, Nicholas T.
author_facet Lagundžin, Dragana
Krieger, Kimiko L.
Law, Henry C.-H.
Woods, Nicholas T.
author_sort Lagundžin, Dragana
collection PubMed
description 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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spelling pubmed-89209162022-03-16 An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry Lagundžin, Dragana Krieger, Kimiko L. Law, Henry C.-H. Woods, Nicholas T. STAR Protoc Protocol 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). Elsevier 2022-03-10 /pmc/articles/PMC8920916/ /pubmed/35300004 http://dx.doi.org/10.1016/j.xpro.2022.101234 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Lagundžin, Dragana
Krieger, Kimiko L.
Law, Henry C.-H.
Woods, Nicholas T.
An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
title An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
title_full An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
title_fullStr An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
title_full_unstemmed An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
title_short An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
title_sort optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8920916/
https://www.ncbi.nlm.nih.gov/pubmed/35300004
http://dx.doi.org/10.1016/j.xpro.2022.101234
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