EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K9)

The EpiQuik™ Histone Methyltransferase Activity/Inhibition Kit (H3-K9) is a convenient set of tools that allows the experimenter to measure the activity or inhibition of individual histone methyltransferase that specifically target histone H3 at lysine 9 (H3-K9). The kit is ready-to-use and provides all the essential components needed to carry out a successful HMT activity/inhibition experiment without the need for radioactivity or any special equipment. The kit has the following advantages:

• Very rapid procedure, which can be completed within 3 hours.
• Safe and innovative colorimetric assay without radioactivity, extraction, and chromatography.
• Specific measurement of activity/inhibition of H3-K9 histone methyltransferases.
• Strip microplate format makes the assay flexible: manual or high throughput analysis.
• Extremely simple, reliable, and consistent assay conditions.

Principle & Procedure
The EpiQuik™ Histone Methyltransferase Activity/Inhibition Assay Kit (H3-K9) is designed for measuring HMTs that specifically target histone H3 at lysine 9. In an assay with this kit, the histone substrate is stably captured on the strip wells. HMT enzymes transfer a methyl group to histone H3 substrate from Adomet to methylate the substrate at lysine 9. The methylated histone H3 -K9 can be recognized with a high-affinity antibody. The ratio or amount of methylated H3-K9, which is directly proportional to enzyme activity, can be quantified through a HRP conjugated secondary antibody-color development system. The HMT activity is then calculated based on the amount of methylated H3-K9 converted by the HMTs.

HK1 (10X Wash Buffer)
HK2 (Histone Assay Buffer)
HK3 (Adomet)*
HK4 (Biotinylated Substrate, 25 µg/ml)*
HK5 (HMT Standard, 10 µg/ml)*
HK6 (Capture Antibody, 100 µg/ml)*
HK7 (Detection Antibody, 200 µg/ml)*
HK8 (Developing Solution)
HK9 (Stop Solution)
Control Enzyme (150 µg/ml)*
8-Well Assay Strips (with frame)
User Guide

* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.

1. Can the nuclear extract be increased for the reaction?
Yes, but not more than 6 µl.

3. If the protein concentration extracted is low (0.5 µg/µl), how much extraction can be added in the reaction?
Increase the concentration of nuclear protein during protein extraction (using more sample amount or reduce volume), and then add more sample solution to the reaction (up to 6 µl).

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