染色质免疫共沉淀实验方法

Protocols for Chromatin Immunoprecipitation

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10~3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。

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.Ch IP 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 Ch IP and ultra-low-input micrococcal nuclease-based native ChIP(ULI-NChIP),which can be applied to rare cell populations at the 103 cell level.