Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA.
Department of Biochemistry, Chemistry Institute, University of São Paulo, São Paulo, Brazil.
Methods Mol Biol. 2023;2599:21-31. doi: 10.1007/978-1-0716-2847-8_3.
Chromatin immunoprecipitation (ChIP) has been used over three decades to characterize protein-DNA interactions in cells and tissues. The samples are initially cross-linked with formaldehyde and subjected to lysis with detergents followed by sonication to generate smaller fragments of DNA covalently bound to nuclear proteins. Antibodies against the protein of interest are then used to immunoprecipitate the protein-DNA complex, allowing for the purification of the genomic locations bound to the protein of interest. The DNA can then be analyzed by several techniques such as PCR, quantitative PCR, DNA microarrays, and next-generation sequencing (NGS). ChIP can be used in cell culture systems and animal tissues to provide insights into how the identity of cells and tissues is established, and how transcriptional programs can be disrupted or reactivated in disease states.
染色质免疫沉淀（ChIP）技术已经被用于研究细胞和组织中的蛋白质-DNA 相互作用超过三十年。该方法首先用甲醛交联样本，然后用去污剂裂解细胞，再进行超声处理，以产生与核蛋白共价结合的较小 DNA 片段。接着，用针对目标蛋白的抗体来免疫沉淀蛋白-DNA 复合物，从而实现对与目标蛋白结合的基因组区域的纯化。随后，可以通过几种技术来分析 DNA，如 PCR、定量 PCR、DNA 微阵列和下一代测序（NGS）。ChIP 可用于细胞培养系统和动物组织，以深入了解细胞和组织的身份是如何建立的，以及转录程序在疾病状态下如何被破坏或重新激活。
