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

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-31 DOI:10.1016/j.xpro.2025.103616
Lei Li, May G Akl, Scott B Widenmaier
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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.1.

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从小鼠肝核分离的纯化染色质的体内染色质免疫沉淀方案。
染色质免疫沉淀(ChIP)用于研究转录因子与基因组的结合,但它可能存在问题。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dna -蛋白复合物的方案。我们描述了肝脏分解和交联,DNA-蛋白复合物分离,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。有关本协议使用和执行的完整细节,请参阅Akl等人1。
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来源期刊
STAR Protocols
STAR Protocols Biochemistry, Genetics and Molecular Biology-General Biochemistry, Genetics and Molecular Biology
CiteScore
2.00
自引率
0.00%
发文量
789
审稿时长
10 weeks
期刊介绍:
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