The EpiQuik™ Histone Methyltransferase Activity/Inhibition Assay Kit (H3-K27) 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 27 (H3-K27). 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.

WHY CHOOSE THE EPIQUIK™ HISTONE METHYLTRANSFERASE ACTIVITY/INHIBITION ASSAY KIT (H3-K27)?

• 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-K27 histone methyltransferases.
• Strip microplate format makes the assay flexible: manual or high throughput analysis.
• Extremely simple, reliable, and consistent assay conditions.

The EpiQuik™ Histone Methyltransferase Activity/Inhibition Assay Kit (H3-K27) is designed for measuring HMTs that specifically target histone H3 at lysine 27. In an assay with this kit, the histone substrate is stably captured on the strip wells through biotin-streptavidin binding. HMT enzymes transfer a methyl group to histone H3 substrate from Adomet to methylate the substrate at lysine 4. The methylated histone H3-K27 can be recognized with a high-affinity antibody. The ratio or amount of methylated H3-K27, 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-K27 converted by the HMTs.

SCHEMATIC PROCEDURE:
	Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit and H3-K27 specific histone methyltrasferase activity was measured.

HE1 (10X Wash Buffer)
HE2 (Histone Assay Buffer)
HE3 (Adomet)*
HE4 (Biotinylated Substrate, 25 µg/ml)*
HE5 (HMT Standard, 10 µg/ml)*
HE6 (Capture Antibody, 100 µg/ml)*
HE7 (Detection Antibody, 100 µg/ml)*
HE8 (Developing Solution)
HE9 (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.

[User Guide]* *Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

[Material Safety Data Sheet]

Rugg-Gunn, PJ. et. al. (June 2010). Distinct histone modifications in stem cell lines and tissue lineages from the early mouse embryo. Proc Natl Acad Sci USA. 107(24):10783-90. PubMed Abstract

Nakade, K. et. al. (April 2009). JDP2 (Jun Dimerization Protein 2)-deficient mouse embryonic fibroblasts are resistant to replicative senescence. The Journal of Biological Chemistry. 284(16): 10808-17. PubMed Abstract