Read Jordan E
Department of Biochemical Pharmacology, William Harvey Research Institute, Queen Mary University of London, London, EC1M 6BQ, UK.
Methods Mol Biol. 2017;1651:23-32. doi: 10.1007/978-1-4939-7223-4_3.
Chromatin immunoprecipitation (ChIP) has become a widely used methodology for assessment of protein/DNA interactions. The technique allows the analysis of direct binding of transcription factors to gene promoters, identification of histone modifications, and localization of DNA modifying enzymes. Antibodies conjugated to agarose beads can be utilized to immunoprecipitate specific proteins, cross-linked to sheared chromatin regions to which they are bound endogenously. With downstream applications including quantitative real-time polymerase chain reaction (qRT-PCR), genome-wide sequencing (ChIP-seq), microarray analysis (ChIP-chip), and mass spectrometry (ChIP-MS), the technique enables comprehensive assessment of protein/DNA interactions. Here I describe ChIP, followed by qRT-PCR, to assess direct binding of a single protein to multiple predicted binding sites within a gene promoter.
染色质免疫沉淀法（ChIP）已成为评估蛋白质/DNA相互作用的一种广泛应用的方法。该技术可用于分析转录因子与基因启动子的直接结合、鉴定组蛋白修饰以及定位DNA修饰酶。与琼脂糖珠偶联的抗体可用于免疫沉淀特定蛋白质，这些蛋白质与内源性结合的剪切染色质区域交联。通过包括定量实时聚合酶链反应（qRT-PCR）、全基因组测序（ChIP-seq）、微阵列分析（ChIP-chip）和质谱分析（ChIP-MS）在内的下游应用，该技术能够全面评估蛋白质/DNA相互作用。在此，我将描述ChIP，随后介绍qRT-PCR，以评估单个蛋白质与基因启动子内多个预测结合位点的直接结合。
