Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen
Summary: Protein synthesis could control spermatogenic cell fate transitions. Here, we present a protocol for visualization and quantification of newly synthesized proteins by click-chemistry-based immunofluorescence within specific spermatogenic cell types in the mice testicular lumen. We detail th...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672500067X |
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| author | Dingfeng Zou Kai Li Yan Lu Wei Yan Wei Song |
| author_facet | Dingfeng Zou Kai Li Yan Lu Wei Yan Wei Song |
| author_sort | Dingfeng Zou |
| collection | DOAJ |
| description | Summary: Protein synthesis could control spermatogenic cell fate transitions. Here, we present a protocol for visualization and quantification of newly synthesized proteins by click-chemistry-based immunofluorescence within specific spermatogenic cell types in the mice testicular lumen. We detail the processes for O-propargyl-puromycin (OPP) incorporation, antibody incubation, confocal microscope imaging, and subsequent quantification methods. This protocol is not limited to spermatogenic cells and can be adapted to investigate protein synthesis in other testicular cell types and various tissue-specific cell populations.For complete details on the use and execution of this protocol, please refer to Zou et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-0d9fca4577444d77adb78be6b2a24ff2 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-0d9fca4577444d77adb78be6b2a24ff22025-08-20T01:47:28ZengElsevierSTAR Protocols2666-16672025-03-016110366110.1016/j.xpro.2025.103661Protocol for identifying protein synthesis activity in specific cell types of the testicular lumenDingfeng Zou0Kai Li1Yan Lu2Wei Yan3Wei Song4Department of Biochemistry and Molecular Biology, State Key Laboratory of Common Mechanism Research for Major Diseases, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China; Corresponding authorDepartment of Biochemistry and Molecular Biology, State Key Laboratory of Common Mechanism Research for Major Diseases, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, ChinaDepartment of Biochemistry and Molecular Biology, State Key Laboratory of Common Mechanism Research for Major Diseases, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, ChinaThe Lundquist Institute for Biomedical Innovation at Harbor-UCLA Medical Center, Torrance, CA 90502, USA; Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA; Corresponding authorDepartment of Biochemistry and Molecular Biology, State Key Laboratory of Common Mechanism Research for Major Diseases, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China; Corresponding authorSummary: Protein synthesis could control spermatogenic cell fate transitions. Here, we present a protocol for visualization and quantification of newly synthesized proteins by click-chemistry-based immunofluorescence within specific spermatogenic cell types in the mice testicular lumen. We detail the processes for O-propargyl-puromycin (OPP) incorporation, antibody incubation, confocal microscope imaging, and subsequent quantification methods. This protocol is not limited to spermatogenic cells and can be adapted to investigate protein synthesis in other testicular cell types and various tissue-specific cell populations.For complete details on the use and execution of this protocol, please refer to Zou et al.1 : 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/S266616672500067XCell BiologyDevelopmental biologyMolecular Biology |
| spellingShingle | Dingfeng Zou Kai Li Yan Lu Wei Yan Wei Song Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen STAR Protocols Cell Biology Developmental biology Molecular Biology |
| title | Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen |
| title_full | Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen |
| title_fullStr | Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen |
| title_full_unstemmed | Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen |
| title_short | Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen |
| title_sort | protocol for identifying protein synthesis activity in specific cell types of the testicular lumen |
| topic | Cell Biology Developmental biology Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S266616672500067X |
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