Beischlag Timothy V, Prefontaine Gratien G, Hankinson Oliver
Faculty of Health Sciences, Simon Fraser University, 8888 University Drive, Burnaby, BC, Canada, V5A 1S6.
Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
Methods Mol Biol. 2018;1689:103-112. doi: 10.1007/978-1-4939-7380-4_9.
Chromatin immunoprecipitation (ChIP) exploits the specific interactions between DNA and DNA-associated proteins. It can be used to examine a wide range of experimental parameters. A number of proteins bound at the same genomic location can identify a multi-protein chromatin complex where several proteins work together to regulate gene transcription or chromatin configuration. In many instances, this can be achieved using sequential ChIP; or simply, ChIP-re-ChIP. Whether it is for the examination of specific transcriptional or epigenetic regulators, or for the identification of cistromes, the ability to perform a sequential ChIP adds a higher level of power and definition to these analyses. In this chapter, we describe a simple and reliable method for the sequential ChIP assay.
染色质免疫沉淀(ChIP)利用DNA与DNA相关蛋白之间的特异性相互作用。它可用于检测多种实验参数。在同一基因组位置结合的多种蛋白质能够鉴定出一种多蛋白染色质复合物,其中几种蛋白质共同作用以调节基因转录或染色质构型。在许多情况下,这可以通过连续ChIP来实现;或者简单地说,就是ChIP再ChIP。无论是用于检测特定的转录或表观遗传调节因子,还是用于鉴定顺反组,进行连续ChIP的能力都为这些分析增添了更高水平的效能和清晰度。在本章中,我们描述了一种用于连续ChIP分析的简单可靠方法。