University of Massachusetts Medical School Faculty Publications
Title
Chromatin immunoprecipitation assay of brain tissues using Percoll gradient-purified nuclei
UMMS Affiliation
Department of Microbiology and Physiological Systems
Publication Date
2013
Document Type
Book Chapter
Disciplines
Laboratory and Basic Science Research | Molecular and Cellular Neuroscience | Molecular Biology | Neuroscience and Neurobiology
Abstract
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.
DOI of Published Version
10.1007/978-1-62703-444-9_19
Source
Methods Mol Biol. 2013;1018:199-209. doi: 10.1007/978-1-62703-444-9_19. Link to article on publisher's site
Related Resources
Journal/Book/Conference Title
Methods in molecular biology (Clifton, N.J.)
PubMed ID
23681630
Repository Citation
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.
https://escholarship.umassmed.edu/faculty_pubs/232