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从小鼠肝细胞核中分离出的纯化染色质进行体内染色质免疫沉淀的实验方案。

Protocol for in vivo chromatin immunoprecipitation on purified chromatin isolated from mouse liver nuclei.

作者信息

Li Lei, Akl May G, Widenmaier Scott B

机构信息

Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada.

Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103616. doi: 10.1016/j.xpro.2025.103616. Epub 2025 Jan 31.

Abstract

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..

摘要

染色质免疫沉淀(ChIP)用于研究转录因子与基因组的结合,但可能存在问题。我们提出了一种在染色质剪切之前从小鼠肝脏中分离固定的DNA-蛋白质复合物的方案。我们描述了肝脏解离和交联、DNA-蛋白质复合物分离、染色质剪切和质量控制分析的步骤,以及ChIP、DNA纯化和ChIP分析的程序。该方案使用商业抗体产生高质量的样品。有关本方案使用和执行的完整详细信息,请参考阿克尔等人的研究。

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