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EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K27)

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Suggested Workflow
Nuclear Protein Extraction
 
 
HMT Assay
 
Schematic procedure for using the EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K27).
Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit and H3-K27 specific histone methyltrasferase activity 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-3005-4848 assays $327.00 
P-3005-9696 assays $551.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

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. 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-K27 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-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.

 
Fig. 1. Schematic procedure for using the EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K27).

Fig. 2. Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit and H3-K27 specific histone methyltrasferase activity was measured.

Product Components

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 & 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

Kowluru RA et. al. (January 2017). Role of oxidative stress in epigenetic modification of MMP-9 promoter in the development of diabetic retinopathy. Graefes Arch Clin Exp Ophthalmol.

Cechinel LR et. al. (October 2016). Treadmill exercise induces age and protocol-dependent epigenetic changes in prefrontal cortex of Wistar rats. Behav Brain Res. 313:82-7.

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

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.

Zhang J et. al. (September 2015). Disruption of KMT2D perturbs germinal center B cell development and promotes lymphomagenesis. Nat Med.

Wan J et. al. (April 2015). PCAF-primed EZH2 acetylation regulates its stability and promotes lung adenocarcinoma progression. Nucleic Acids Res. 43(7):3591-604.

De la Cruz-Hernandez E et. al. (February 2015). Ribavirin as a tri-targeted antitumor repositioned drug. Oncol Rep.

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

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.

Malmgren S et. al. (April 2013). Coordinate changes in histone modifications, mRNA levels, and metabolite profiles in clonal INS-1 832/13 β-cells accompany functional adaptations to lipotoxicity. J Biol Chem. 288(17):11973-87.

Latrasse D et. al. (March 2013). Dual function of MIPS1 as a metabolic enzyme and transcriptional regulator. Nucleic Acids Res. 41(5):2907-17.

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.

Batra V et. al. (March 2012). Interaction between γ-radiation and dietary folate starvation metabolically reprograms global hepatic histone H3 methylation at lysine 4 and lysine 27 residues. Food Chem Toxicol. 50(3-4):464-72.

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 U S A. 107(24):10783-90.

Nakade K et. al. (April 2009). JDP2 (Jun Dimerization Protein 2)-deficient mouse embryonic fibroblasts are resistant to replicative senescence. J Biol Chem. 284(16):10808-17.

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