The EpiQuik™ Histone Demethylase (H3-K9 Specific) Activity/Inhibition Fast Assay Kit is a convenient set of tools that allows the experimenter to measure activity/inhibition of H3-K9 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-K9 specific) activity/inhibition from a broad range of species including mammalian cells/tissues, plants, and bacteria. The kit has the following advantages:

• Fast one-step assay, which can be finished within 1.5 hours.
• Innovative fluorescent assay without the need for radioactivity, extraction, and chromatography.
• Measurement of HDM (H3-K9 specific) activity and inhibition through quantifying by-product of the enzyme reaction, which make the assay highly sensitive.
• 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-K9 Specific) Activity/Inhibition Fast Assay Kit is designed for measuring total histone demethylase (H3-K9 specific) activity/inhibition. In the assay with this kit, the unique methylated histone H3-K9 substrate is incubated with nuclear extracts or purified enzymes. Formaldehyde generated from the enzyme-substrate reaction reacts with a detection reagent to form a fluorescent substance. The ratio or amount of the formaldehyde, which is proportional to HDM enzyme activity, can then be fluorometrically quantified.

 
Fig. 1. Schematic procedure for using the EpiQuik Histone Demethylase (H3K9 Specific) Activity/Inhibition Fast Assay Kit.

Fig. 2. HDM (H3-K9 specific) activity measured from HeLa cells using the EpiQuik™ Histone Demethylase (H3-K9 Specific) Activity/Inhibition Fast Assay Kit.