An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins
Summary: Ubiquitination modification is an important post-translational modification that regulates the stability and function of proteins. Here, we present a protocol to detect the K27-linked polyubiquitination of exogenous and endogenous mitochondrial antiviral signaling protein. We describe steps...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723006172 |
| _version_ | 1797654726497533952 |
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| author | Xinyu Li Wei Jiang Min Fang |
| author_facet | Xinyu Li Wei Jiang Min Fang |
| author_sort | Xinyu Li |
| collection | DOAJ |
| description | Summary: Ubiquitination modification is an important post-translational modification that regulates the stability and function of proteins. Here, we present a protocol to detect the K27-linked polyubiquitination of exogenous and endogenous mitochondrial antiviral signaling protein. We describe steps for detecting ubiquitination of exogenous protein, transfecting the encoding plasmid of the protein, and immunoprecipitating the target protein with an antibody. We then detail procedures for detecting ubiquitin of the target protein by western blot. This protocol applies to other proteins of interest.For complete details on the use and execution of this protocol, please refer to Jiang et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-11T17:03:41Z |
| format | Article |
| id | doaj.art-d7a203a780844f6c8a663c2e02b51dce |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-11T17:03:41Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-d7a203a780844f6c8a663c2e02b51dce2023-10-20T06:48:42ZengElsevierSTAR Protocols2666-16672023-12-0144102650An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteinsXinyu Li0Wei Jiang1Min Fang2CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, ChinaCAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, ChinaCAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; International College, University of Chinese Academy of Sciences, Beijing 100049, China; School of Life Sciences, Henan University, Kaifeng, Henan 475004, China; Corresponding authorSummary: Ubiquitination modification is an important post-translational modification that regulates the stability and function of proteins. Here, we present a protocol to detect the K27-linked polyubiquitination of exogenous and endogenous mitochondrial antiviral signaling protein. We describe steps for detecting ubiquitination of exogenous protein, transfecting the encoding plasmid of the protein, and immunoprecipitating the target protein with an antibody. We then detail procedures for detecting ubiquitin of the target protein by western blot. This protocol applies to other proteins of interest.For complete details on the use and execution of this protocol, please refer to Jiang et al. (2023).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/S2666166723006172Cell-Based AssaysCell Separation/FractionationImmunologyMolecular Biology |
| spellingShingle | Xinyu Li Wei Jiang Min Fang An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins STAR Protocols Cell-Based Assays Cell Separation/Fractionation Immunology Molecular Biology |
| title | An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins |
| title_full | An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins |
| title_fullStr | An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins |
| title_full_unstemmed | An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins |
| title_short | An optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins |
| title_sort | optimized protocol to detect ubiquitination modification of exogenous or endogenous proteins |
| topic | Cell-Based Assays Cell Separation/Fractionation Immunology Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723006172 |
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