Description
A new application note from Porvair Sciences details a protocol for its Chromatrap® ChIP assay kit that has been developed to enrich epigenetic marks from primary human tissue cells.Chromatrap® Pro-A, the novel solid-based matrix for chromatin immunoprecipitation assays (ChIP) has been successfully used for isolation of high quality chromatin from difficult biopsy material. Focusing on an application to precipitate epigenetic marks from primary endometrial stromal cells - results show high abundant epigenetic marks from 1 million cells isolated from a tissue biopsy, with selective amplification of positive gene targets relative to negative IgG and negative gene loci.
Not only is Chromatrap proven to produce positive amplification of histone regulatory marks at specific gene loci but also the novel ChIP assay kit is shown to be compatible with human tissue.
Chromatrap® uses a solid phase porous polymer functionalised with protein A, allowing the chromatin capture to be more efficient than bead based methods. The Chromatrap spin column approach offers significant advantages compared to methods based on sepharose or magnetic beads, which involve many steps of separation, pipetting and re-suspension. Independent tests with Chromatrap demonstrate levels of DNA pull down up to 25 times higher than with traditional bead based procedures, excellent DNA enrichment with signal-to-noise ratios typically 2 to 3 times better than competing procedures and top performance even with samples as small as 50ng or as much as 3000ng chromatin per immunoprecipitation.
