Li Lei, Akl May G, Widenmaier Scott B
Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada.
Department of Anatomy, Physiology, and Pharmacology, University of Saskatchewan, Saskatoon, SK, Canada.
STAR Protoc. 2025 Mar 21;6(1):103616. doi: 10.1016/j.xpro.2025.103616. Epub 2025 Jan 31.
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..
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。
