Malik Victoria Adelheid, Di Benedetto Barbara, Jakovcevski Mira
Department of Psychiatry and Psychotherapy, Faculty of Medicine, University of Regensburg, Regensburg, Germany.
Regensburg Center of Neuroscience, University of Regensburg, Regensburg, Germany.
Methods Mol Biol. 2019;1938:219-229. doi: 10.1007/978-1-4939-9068-9_15.
Chromatin immunoprecipitation (ChIP) in conjunction with qPCR or next generation sequencing (ChIP-seq) is used to detect protein-DNA interaction. Typically, DNA bound to a protein of interest is captured with an antibody against this protein, and DNA is then purified from DNA-protein complexes. Here, we describe a native Chromatin immunoprecipitation (N-ChIP) approach which is an efficient ChIP method with high resolution for histone modifications and a number of transcription factors. This protocol has been tailored for cultured primary rat astrocytes, and we included the preparation of astrocytic cell cultures in this protocol.
染色质免疫沉淀（ChIP）结合定量聚合酶链反应（qPCR）或下一代测序（ChIP-seq）用于检测蛋白质与DNA的相互作用。通常，用针对目标蛋白质的抗体捕获与该蛋白质结合的DNA，然后从DNA-蛋白质复合物中纯化DNA。在此，我们描述了一种天然染色质免疫沉淀（N-ChIP）方法，它是一种高效的ChIP方法，对组蛋白修饰和许多转录因子具有高分辨率。该方案是针对原代培养的大鼠星形胶质细胞定制的，我们在本方案中纳入了星形胶质细胞培养物的制备。
