The EpiQuik™ Histone Demethylase (H3-K4 Specific) Activity/Inhibition Assay Kit is a convenient set of tools that allows the experimenter to measure activity/inhibition of H3-K4 specific histone demethylases using cell/ tissue extracts. The kit is ready-to-use and provides all the essential components needed for measuring histone demethylase (H3-K4 Specific) activity/inhibition from a broad range of species including mammalian cells/tissues, plants, and bacteria. The kit has the following advantages:
- Fast procedure, which can be finished within 3 hours.
- Innovative fluorescent assay with no use of radioactivity, extraction, and chromatography.
- Direct measurement of HDM (H3-K4 specific) activity and inhibition without interference by thiol-containing chemicals such as DTT, GSH, and 2-mercaptoethanol.
- Strip microplate format makes the assay flexible: manual or high throughput analysis.
- Simple, reliable, and consistent assay conditions.
Principle & Procedure
The EpiQuik™ Histone Demethylase (H3-K4 specific) Activity/Inhibition Assay Kit is designed for measuring total histone demetylase (H3-K4 Specific) activity/inhibition. In the assay with this kit, the unique di-methylated histone H3-K4 substrate is stably captured on the strip wells. Active HDMs bind to and demethylate histone H3-K4 substrate. The remaining un-demethylated substrate can be recognized with a high affinity anti-methylated histone H3-K4 antibody. The ratio or amount of the un-demethylated histone, which is inversely proportional to HDM enzyme activity, can then be fluorometrically quantified.
Fig. 1. Schematic procedure for using the EpiQuik Histone Demethylase (H3K4 Specific) Activity/Inhibition Assay Kit.
Fig. 2. HDM (H3-K4 specific) activity measured from Hela cell extracts.