Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei
Summary: Chromatin immunoprecipitation (ChIP) is used to investigate genome binding by transcription factors, but it can be problematic. We present a protocol to isolate fixed DNA-protein complexes from mouse liver prior to chromatin shearing. We describe steps for liver disaggregation and cross-lin...
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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/S266616672500022X |
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| author | Lei Li May G. Akl Scott B. Widenmaier |
| author_facet | Lei Li May G. Akl Scott B. Widenmaier |
| author_sort | Lei Li |
| collection | DOAJ |
| description | Summary: Chromatin immunoprecipitation (ChIP) is used to investigate genome binding by transcription factors, but it can be problematic. We present a protocol to isolate fixed DNA-protein complexes from mouse liver prior to chromatin shearing. We describe steps for liver disaggregation and cross-linking, DNA-protein complex isolation, chromatin shearing, and quality control analysis as well as procedures for ChIP, DNA purification, and ChIP analysis. This protocol yields high-quality samples using commercial antibodies.For complete details on the use and execution of this protocol, please refer to Akl 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-c26759ab2fa44ae0bd22cdf2cea2b16d |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-c26759ab2fa44ae0bd22cdf2cea2b16d2025-02-02T05:29:21ZengElsevierSTAR Protocols2666-16672025-03-0161103616Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nucleiLei Li0May G. Akl1Scott B. Widenmaier2Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, CanadaDepartment of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, CanadaDepartment of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada; Corresponding authorSummary: Chromatin immunoprecipitation (ChIP) is used to investigate genome binding by transcription factors, but it can be problematic. We present a protocol to isolate fixed DNA-protein complexes from mouse liver prior to chromatin shearing. We describe steps for liver disaggregation and cross-linking, DNA-protein complex isolation, chromatin shearing, and quality control analysis as well as procedures for ChIP, DNA purification, and ChIP analysis. This protocol yields high-quality samples using commercial antibodies.For complete details on the use and execution of this protocol, please refer to Akl 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/S266616672500022Xmolecular biologyantibodychromatin immunoprecipitationChIP |
| spellingShingle | Lei Li May G. Akl Scott B. Widenmaier Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei STAR Protocols molecular biology antibody chromatin immunoprecipitation ChIP |
| title | Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei |
| title_full | Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei |
| title_fullStr | Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei |
| title_full_unstemmed | Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei |
| title_short | Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei |
| title_sort | protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei |
| topic | molecular biology antibody chromatin immunoprecipitation ChIP |
| url | http://www.sciencedirect.com/science/article/pii/S266616672500022X |
| work_keys_str_mv | AT leili protocolforinvivochromatinimmunoprecipitationonpurifiedchromatinisolatedfrommouselivernuclei AT maygakl protocolforinvivochromatinimmunoprecipitationonpurifiedchromatinisolatedfrommouselivernuclei AT scottbwidenmaier protocolforinvivochromatinimmunoprecipitationonpurifiedchromatinisolatedfrommouselivernuclei |