Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis

Here, we describe a protocol for extraction and tandem immunoprecipitation of the cytoplasmic and nuclear proteins from mouse testis for mass spectrometry. This protocol has been applied to knockin mice that express a meiotic protein of interest tagged with 3xFLAG-HA in the testis. The protocol is o...

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Detalles Bibliográficos
Autores principales: Tani, Naoki, Tanno, Nobuhiro, Ishiguro, Kei-ichiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9201064/
https://www.ncbi.nlm.nih.gov/pubmed/35719267
http://dx.doi.org/10.1016/j.xpro.2022.101452
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author Tani, Naoki
Tanno, Nobuhiro
Ishiguro, Kei-ichiro
author_facet Tani, Naoki
Tanno, Nobuhiro
Ishiguro, Kei-ichiro
author_sort Tani, Naoki
collection PubMed
description Here, we describe a protocol for extraction and tandem immunoprecipitation of the cytoplasmic and nuclear proteins from mouse testis for mass spectrometry. This protocol has been applied to knockin mice that express a meiotic protein of interest tagged with 3xFLAG-HA in the testis. The protocol is optimized for salt extraction of cytoplasmic and nuclear proteins from mouse frozen testes and thus can be used for a variety of proteins. For complete details on the use and execution of this protocol, please refer to Ishiguro et al. (2020) and Tanno et al. (2022).
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spelling pubmed-92010642022-06-17 Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis Tani, Naoki Tanno, Nobuhiro Ishiguro, Kei-ichiro STAR Protoc Protocol Here, we describe a protocol for extraction and tandem immunoprecipitation of the cytoplasmic and nuclear proteins from mouse testis for mass spectrometry. This protocol has been applied to knockin mice that express a meiotic protein of interest tagged with 3xFLAG-HA in the testis. The protocol is optimized for salt extraction of cytoplasmic and nuclear proteins from mouse frozen testes and thus can be used for a variety of proteins. For complete details on the use and execution of this protocol, please refer to Ishiguro et al. (2020) and Tanno et al. (2022). Elsevier 2022-06-13 /pmc/articles/PMC9201064/ /pubmed/35719267 http://dx.doi.org/10.1016/j.xpro.2022.101452 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
Tani, Naoki
Tanno, Nobuhiro
Ishiguro, Kei-ichiro
Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
title Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
title_full Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
title_fullStr Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
title_full_unstemmed Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
title_short Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
title_sort tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for ms analysis
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9201064/
https://www.ncbi.nlm.nih.gov/pubmed/35719267
http://dx.doi.org/10.1016/j.xpro.2022.101452
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AT tannonobuhiro tandemimmunopurificationofaffinitytaggedproteinsfrommousetestisextractsformsanalysis
AT ishigurokeiichiro tandemimmunopurificationofaffinitytaggedproteinsfrommousetestisextractsformsanalysis

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