药物化学

Native Chromatin Immunoprecipitation from Brain Tissue Using Magnetic Beads

Abstract

Lene Lundgaard Donovan and Jacek Lichota

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.