Description

Porvair Sciences has announced an application note describing the excellent results achievable from a new 96-well format version of its ChromaTrap  chromatin immunoprecipitation (ChIP) assay technology.

Developments in the field of epigenetics are rapidly expanding as researchers elucidate the principles governing gene regulation through the in vivo dynamic binding of proteins to DNA. Key to this is the need to analyse large numbers of high and low abundant regulatory mechanisms to systematically map protein-DNA interactions following pathogen/disease stimulus.

Chromatin immunoprecipitation (ChIP) assays are used to study these associations and have been historically limited to large sample numbers, onerous washing steps resulting in valuable sample losses and low signal to noise coupled with poor reproducibility. This new application note details the use of Chromatrap®96 Pro-A, a 96-well plate, each well of which contains a solid phase rigid porous polymer, BioVyonTM, to which Protein A is chemically bound, allowing up to 96 ChIP assays to be run in parallel on the same plate.

To determine ease of use, experimental design flexibility and reproducibility in screening multiple epigenetic targets with Chromatrap®96, the binding efficiency and occupancy of common epigenetic marks (H3, H4, H3K4me3 and RNA pol II) on three gene targets of interest (GAPDH, beta-globin, and PABPC1) in three human chromatin samples (HepG2, HeLa and K562) were performed simultaneously on the 96-well plate and analysed using qPCR. The application note describes in detail the analytical methodology, data interpretation and statistical analysis, as well as the benefits of using the new Chromatrap®96 high throughput ChIP assay plate which include multiple target screening, flexible experimental design, robust reproducibility, high data quality and automated liquid handling potential.