Epigentek has introduced a method to directly measure histone demethylase (HDMT) activity/inhibition via a straightforward detection of HDMT-converted products rather than by-products.
This method is incorporated into commercially available kits under the Epigenase brand for histone demethylase activity/inhibition assay kit series, which provide ultra-sensitive assays of LSD1, JMJD2, and JARID demethylase activity or inhibition.
The discovery of histone methylases demonstrates that histone methylation is not a permanent modification, but rather a more dynamic process.
LSD1, JMJD2 and JARID are major histone demethylases and have been found to be essential in cancer development.
Detection of activity and inhibition of these enzymes would be important in elucidating epigenetic mechanisms of gene function regulation and in cancer drug development, thereby benefiting cancer diagnostics and therapeutics.
Currently used methods are based on the measurement of H2O2/formaldehyde release, a by-product of HDMT enzymatic reaction and have significant weaknesses: large amounts (at ug level) of substrates and enzymes are required; nuclear extracts from cell/tissues cannot be used; redox-sensitive HDMT inhibitors are not suitable for testing with these methods; high interference by SDS, DMSO, ions, and thiol-containing chemicals, which are often found in enzyme solutions, tested compound solvents, and assay buffers; and less accuracy than a direct measurement approach of HDMT-converted demethylated products.
In contrast, the Epigenase method is said to address all of the aforementioned shortcomings.
The immunoaffinity based-Epigenase method is able to directly detect HDMT-converted end-products, rather than by-products.
Therefore it eliminates any assay interference caused by a variety of chemicals and detergents, and has a much higher sensitivity (up to 2,000 times) and specificity with the ability to measure HDMT activity/inhibition using either purified enzymes or cell/tissue extracts.
The kits based on the Epigenase method allows entire assays to be completed within just 3.5 hours in a 96 stripwell microplate format, which would be particularly useful for high-throughput screening in discovering inhibitors of HDMTs as potential therapeutics agents.
