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EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K9)

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Suggested Workflow
Nuclear Protein Extraction
 
 
HMT Assay
 
Schematic procedure for using the EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K9).
Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit and H3-K9 specific histone methyltrasferase activity (G9a) was measured.
Input Type: Nuclear Extracts, Purified Enzyme
Research Area: Histone Methylation
Target Application: Activity Measurement
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-3003-148 assays $327.00 
P-3003-296 assays $551.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

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.

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

Fig. 2. Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit and H3-K9 specific histone methyltrasferase activity (G9a) was measured.

Product Components

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.

Frequently Asked Q's

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).

User Guide & MSDS

[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]
Product Citations

Xue Y et. al. (July 2017). RIZ1 and histone methylation status in pituitary adenomas. Tumour Biol. 39(7):1010428317711794.

Guan M et. al. (June 2017). Fatty acid synthase reprograms the epigenome in uterine leiomyosarcomas. PLoS One. 12(6):e0179692.

Kenji Ishimoto et. al. (October 2016). Ubiquitination of Lysine 867 of the Human SETDB1 Protein Upregulates Its Histone H3 Lysine 9 (H3K9) Methyltransferase Activity. PLOS One.

NavakauskienÄ— et. al. (May 2016). Histone demethylating agents as potential S-adenosyl-L-methionine-competitors Med Chem Comm.

Liu J et. al. (January 2016). Clemastine Enhances Myelination in the Prefrontal Cortex and Rescues Behavioral Changes in Socially Isolated Mice. J Neurosci. 36(3):957-62.

Nizialek EA et. al. (December 2015). Cancer-predisposition gene KLLN maintains pericentric H3K9 trimethylation protecting genomic stability. Nucleic Acids Res.

Veazey KJ et. al. (September 2015). Dose-dependent alcohol-induced alterations in chromatin structure persist beyond the window of exposure and correlate with fetal alcohol syndrome birth defects. Epigenetics Chromatin. 8:39.

Cheedipudi S et. al. (July 2015). A fine balance: epigenetic control of cellular quiescence by the tumor suppressor PRDM2/RIZ at a bivalent domain in the cyclin a gene. Nucleic Acids Res. 43(13):6236-56.

Liu J et. al. (January 2015). Chromatin landscape defined by repressive histone methylation during oligodendrocyte differentiation. J Neurosci. 35(1):352-65.

Austermann J et. al. (December 2014). Alarmins MRP8 and MRP14 Induce Stress Tolerance in Phagocytes under Sterile Inflammatory Conditions. Cell Rep. 9(6):2112-23.

Morishita M et. al. (December 2014). In vitro histone lysine methylation by NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L. BMC Struct Biol. 14(1):25.

Sahdeo S et. al. (August 2014). Dyclonine rescues frataxin deficiency in animal models and buccal cells of patients with Friedreich's ataxia. Hum Mol Genet.

Li B et. al. (July 2013). Identification and characterization of the Spodoptera Su(var) 3-9 histone H3K9 trimethyltransferase and its effect in AcMNPV infection. PLoS One. 8(7):e69442.

Han YC et. al. (June 2013). Metallothionein 1 h tumour suppressor activity in prostate cancer is mediated by euchromatin methyltransferase 1. J Pathol. 230(2):184-93.

Horrillo A et. al. (April 2013). Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment. Cell Death Dis. 4:e570.

Dong C et. al. (March 2013). Interaction with Suv39H1 is critical for Snail-mediated E-cadherin repression in breast cancer. Oncogene. 32(11):1351-62.

Li Y et. al. (January 2013). Epigenetic regulation of multiple tumor-related genes leads to suppression of breast tumorigenesis by dietary genistein. PLoS One. 8(1):e54369.

Zou Y et. al. (July 2012). Effect of phenylhexyl isothiocyanate on aberrant histone H3 methylation in primary human acute leukemia. J Hematol Oncol. 5:36.

Dong C et. al. (April 2012). G9a interacts with Snail and is critical for Snail-mediated E-cadherin repression in human breast cancer. J Clin Invest. 122(4):1469-86.

Candelaria M et. al. (March 2012). DNA methylation-independent reversion of gemcitabine resistance by hydralazine in cervical cancer cells. PLoS One. 7(3):e29181.

Chestnut BA et. al. (November 2011). Epigenetic regulation of motor neuron cell death through DNA methylation. J Neurosci. 31(46):16619-36.

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