Native Chromatin Immunoprecipitation (N-ChIP) in Primary Cortical Rat Astrocytes
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.
Key wordsChIP-sequencing (ChIP-seq) Histone H3 lysine 4 trimethylation (H3K4me3) Native chromatin immunoprecipitation (N-ChIP) Astrocytes
The work of Dr. Mira Jakovcevski (MJ) was supported by a NARSAD Young Investigator Grant (#22809) from the Brain and Behavior Research Foundation. MJ is an “Attias Family Foundation Investigator.” The authors thank Dr. Tobias Straub for analysis of H3K4me3 sequencing data. The work of Dr. Barbara Di Benedetto (BDB) was supported by intramural funding from the University of Regensburg, by the German Federal Ministry of Education and Research (BMBF Grant 01EE1401A), and by the German Research Council (DFG GRK2174).
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