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.
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Ding, B., Kilpatrick, D.L. (2013). Chromatin Immunoprecipitation Assay of Brain Tissues Using Percoll Gradient-Purified Nuclei. In: Zhou, R., Mei, L. (eds) Neural Development. Methods in Molecular Biology, vol 1018. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-444-9_19
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DOI: https://doi.org/10.1007/978-1-62703-444-9_19
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-444-9
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