Chromatin immunoprecipitation assay of brain tissues using Percoll gradient-purified nuclei
Department of Microbiology and Physiological Systems
Laboratory and Basic Science Research | Molecular and Cellular Neuroscience | Molecular Biology | Neuroscience and Neurobiology
Protein-DNA interactions are critical to maintain genome stability, DNA replication, chromosome -segregation and to regulate gene expression. Chromatin immunoprecipitation (ChIP) is a powerful technique to study these interactions within living neurons and nervous tissue. In particular, ChIP analysis of chromatin in which protein-DNA interactions are first fixed in situ provides a valuable approach to identify specific transcription factor-DNA interactions and their regulation in the developing nervous system. Here we describe a procedure utilizing Percoll gradient purification of nuclei from fresh brain tissue pre-fixed with formaldehyde for ChIP analysis. This purification protocol provides an enrichment of neuronal nuclei in high yield. We also illustrate the suitability of chromatin prepared from Percoll-purified brain nuclei for ChIP analysis of regulated transcription factor interactions with neuronal gene promoters.
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Citation: Methods Mol Biol. 2013;1018:199-209. doi: 10.1007/978-1-62703-444-9_19. Link to article on publisher's site
Ding, Baojin and Kilpatrick, Daniel L., "Chromatin immunoprecipitation assay of brain tissues using Percoll gradient-purified nuclei" (2013). University of Massachusetts Medical School Faculty Publications. 232.