Native Chromatin Immunoprecipitation from Brain Tissue Using Magnetic BeadsLene Lundgaard Donovan* and Jacek Lichota
Laboratory of Neurobiology, Department of Health Science and Technology, Aalborg University, Denmark
- *Corresponding Author:
- Lene Lundgaard Donovan
Laboratory of Neurobiology
Department of Health Science and Technology
Aalborg University, Denmark
Tel: +45 27213026
E-mail: [email protected]
Received date: October 22, 2014; Accepted date: November 05, 2014; Published date: November 07, 2014
Citation: Donovan LL, Lichota J (2014) Native Chromatin Immunoprecipitation from Brain Tissue Using Magnetic Beads. Med chem 4:738-741. doi:10.4172/2161-0444.1000223
Copyright: © 2014 Donovan LL, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
We hereby present a protocol for Native Chromatin Immunoprecipitation (NChIP) on brain tissue. Maintaining the chromatin in it’s native state, as opposed to cross-linkage by formaldehyde, and using magnetic beads (instead of sepharose beads) facilitates a very sensitive and specific immunoprecipitation (up to 98% enrichment relative to input). Performing qPCR on acetylated H4 precipitated DNA, we found a 12-fold enrichment of the active actin gene compared to the inactive globin gene. Furthermore, very small inter assay variations were found across individual animals. The high sensitivity and specificity of the present protocol circumvents the need for large tissue samples, which is often a limiting factor when working with brain tissue.