Spin-Column ChIP Assay used with Low-Abundant Targets

Chromatrap® Pro-A, the novel solid-based matrix for chromatin immunoprecipitation assays (ChIP) from Porvair, has been successfully used by researchers to measure low abundant estrogen receptor (ER) transcription factor binding on its target gene loci (XBP1) in the endometrial epithelium.

A recent study by the Reproductive Biology & Cancer Research group at the Institute of Life Science, Swansea University compared the Chromatrap performance to that of the magnetic bead based assay targeted at ERa and the less abundant ERb isoform using a Human endometrial epithelial cell line Ishikawa, grown in the presence of Estradiol and Tamoxifen. Chromatrap was found to be as sensitive in terms of % real signal binding of ERa on the XBP1 promoter as the bead based assay despite using a much lower chromatin input template and concentration (less than half) of ERa antibody. The performance of the Chromatrap ChIP kit was even more robust when used to detect binding of the less abundant ERb isoform on the XBP1 promoter. Full details are available in a newly published Application Note.