Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis
Summary: 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 prot...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672200332X |
| _version_ | 1818470585429131264 |
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| author | Naoki Tani Nobuhiro Tanno Kei-ichiro Ishiguro |
| author_facet | Naoki Tani Nobuhiro Tanno Kei-ichiro Ishiguro |
| author_sort | Naoki Tani |
| collection | DOAJ |
| description | Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-13T21:40:01Z |
| format | Article |
| id | doaj.art-8a501e55ea7e4073a62c5ad0972ab895 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T21:40:01Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-8a501e55ea7e4073a62c5ad0972ab8952022-12-22T02:28:49ZengElsevierSTAR Protocols2666-16672022-06-0132101452Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysisNaoki Tani0Nobuhiro Tanno1Kei-ichiro Ishiguro2Liaison Laboratory Research Promotion Center, IMEG, Kumamoto University, Kumamoto 860-0811, JapanDepartment of Chromosome Biology, Institute of Molecular Embryology and Genetics (IMEG), Kumamoto University, Kumamoto 860-0811, JapanDepartment of Chromosome Biology, Institute of Molecular Embryology and Genetics (IMEG), Kumamoto University, Kumamoto 860-0811, Japan; Corresponding authorSummary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672200332XCell BiologyMolecular BiologyProtein BiochemistryProtein expression and purificationMass Spectrometry |
| spellingShingle | Naoki Tani Nobuhiro Tanno Kei-ichiro Ishiguro Tandem immuno-purification of affinity-tagged proteins from mouse testis extracts for MS analysis STAR Protocols Cell Biology Molecular Biology Protein Biochemistry Protein expression and purification Mass Spectrometry |
| 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 | Cell Biology Molecular Biology Protein Biochemistry Protein expression and purification Mass Spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S266616672200332X |
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