染色质免疫共沉淀实验方法
收稿日期: 2020-05-08
录用日期: 2020-08-13
网络出版日期: 2020-08-26
基金资助
国家自然科学基金(31861130355)
Protocols for Chromatin Immunoprecipitation
Received date: 2020-05-08
Accepted date: 2020-08-13
Online published: 2020-08-26
染色质免疫共沉淀(ChIP)技术是一种检测蛋白质与DNA结合的实验技术。该方法可以先进行样品交联, 然后将蛋白质与DNA复合物进行随机DNA切断, 再借助免疫学方法特异性富集与目的蛋白相结合的DNA片段, 从而检测转录因子等目的蛋白质与DNA的结合情况, 鉴定基因启动子或其它DNA结合位点。该方法同时也可应用于研究基因组特定位点的组蛋白修饰情况。该文介绍了依赖交联固定的常规免疫共沉淀(X-ChIP), 以及适用于103细胞级别微量实验材料的基于微球菌核酸酶非交联免疫共沉淀(ULI-NChIP)具体操作过程和注意事项。
关键词： 染色质免疫共沉淀; 基于微球菌核酸酶非交联免疫共沉淀
王泓力,焦雨铃 . 染色质免疫共沉淀实验方法[J]. 植物学报, 2020 , 55(4) : 475 -480 . DOI: 10.11983/CBB20076
Chromatin immunoprecipitation (ChIP) is a teschnique used to investigate protein-DNA interaction. Briefly, protein and associated DNA are crosslinked, which is optional. Then the DNA-protein complexes are fragmented. By using an appropriate antibody, cross-linked DNA fragments associated with the protein of interest are selectively immunoprecipitated. ChIP is commonly used to identify binding regions of DNA-binding proteins, such as transcription factors. ChIP is also used to analyze histone modification profiles in combination with antibodies against specific histone modifications. Here we describe protocols and related tips for cross-linked ChIP and ultra-low-input micrococcal nuclease-based native ChIP (ULI-NChIP), which can be applied to rare cell populations at the 103 cell level.
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