EpiQuik DNA Demethylase Activity/Inhibition Assay Ultra Kit

For measuring total DNA demethylase activity or inhibition using nuclear extracts from a broad range of species

Product Description

The EpiQuik™ DNA Demethylase Activity/Inhibition Assay Ultra Kit is a complete set of essential components that enables an experimenter to measure total DNA demethylase activity/inhibition. The kit can be used with nuclear extracts from a broad range of species such as mammals and plants, in a variety of forms including, but not limited to, cultured cells and fresh tissues. The EpiQuik™ DNA Demethylase Activity/Inhibition Assay Ultra Kit has the following features:

• Colorimetric assay with easy-to-follow steps for convenience and speed. The entire procedure can be completed within 4 hours.
• Safe and innovative colorimetric assay without the need for radioactivity, extraction, or chromatography.
• An ultra-sensitive detection limit, as low as 1 µg of nuclear extract, is five times more sensitive than the predecessor kit.
• 96 strip-well microplate format allows for either low or high throughput analysis.

Principle & Procedure

In an assay with this kit, the unique methylated DNA substrate is stably captured on the strip wells. Active DNA demethylases bind to and demethylate the DNA substrate. The methylated DNA can be recognized with a high-affinity 5-methylcytosine antibody and the immuno-signal is enhanced with enhancer solution. The ratio or amount of methylated DNA, which is inversely proportional to enzyme activity, can then be colorimetrically quantified through an ELISA-like reaction.

Starting Materials & Input Amount

The amount of nuclear extracts for each assay can be from 2 µg to 20 µg with an optimal range of 5 to 10 µg. We recommend using Epigentek's nuclear extraction kit for optimal results.

Performance Data

Fig.2. Assay control standard was added into the assay wells at different concentrations and then measured with the DNA Demethylase Activity/Inhibition Assay Ultra Kit.