Save $100 on Antibodies Save $100 when you buy $500 in antibodies. Use code E1705ABR. Expires June 30th. » Learn More
Epigentek Home
 
Epigentek Free Shipping

EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Fluorometric)

Share:

For fluorescence-based measurement of DNA methyltransferase activity or inhibition

Citations (5) | Write a Review
Suggested Workflow
Nuclear Protein Extraction
 
 
DNMT Assay
 
Schematic procedure for the EpiQuik™ DNMT Activity/Inhibition Assay Ultra Kit (Fluorometric).
Input Type: Nuclear Extracts, Purified Enzyme
Research Area: DNA Methylation
Target Application: Activity Measurement
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-3010-4848 assays $338.00 
P-3010-9696 assays $549.00 
Availability: Usually Ships in 1 Day or Same day delivery Same Day NY Delivery 
Product Overview

The EpiQuik™ DNMT (DNA Methyltransferase) Activity/Inhibition Assay Ultra Kit (Fluorometric) is a complete set of optimized buffers and reagents that allows the experimenter to fluorometrically measure DNA methyltransferase activity or inhibition at extremely fast speeds on a 96-stripwell microplate. The kit is ready-to-use and provides all the essential components needed to carry out a successful DNMT activity/inhibition experiment without the need for radioactivity or any special equipment. The EpiQuik DNMT Activity/Inhibition Assay Ultra Kit is a further refinement of its popular predecessor kit by enhancing sample signals and significantly minimizing background signals, in addition to being ten times more sensitive. The kit has the following advantages and features:

  • Fluorometric assay with easy-to-follow steps for convenience and speed. The entire procedure can be completed within 3 hours and 45 minutes.
  • Safe and innovative fluorescence assay without radioactivity, extraction, and chromatography.
  • The ultra-sensitive detection limit can be as low as 0.2 µg of nuclear extract or 0.2 ng of purified enzymes, which is ten times better than the predecessor kit.
  • Optimized antibody & enhancer solutions allow high specificity to 5-mC without cross-reactivity to unmethylated cytosine.
  • Strip-well microplate format allows for either low or high throughput analysis.

The EpiQuik Nuclear Extraction Kit has also been optimized for use with this kit.

About DNA Methyltransferases
The addition of methyl groups is carried out by a family of enzymes, DNA methyltransferases (DNMTs or DNA MTases). Chromatin structure in the vicinity of gene promoters also affects DNA methylation and transcriptional activity. DNMT1, DNMT3A, and DNMT3B are required for the establishment and maintenance of DNA methylation patterns. Two additional enzymes, DNMT2/TRDMT1 and DNMT3L, may also have more specialized but related functions. DNMT1 appears to be responsible for maintenance of established patterns of DNA methylation, while DNMT3A and 3B seem to mediate establishment of new or de novo, DNA methylation patterns. DNMT2/TRDMT1 was shown to methylate tRNAAsp at C38 and DNMT3L is found to be a catalytically inactive regulatory factor of DNA methyltransferases, which is essential for the function of DNMT3A and DNMT3B. Diseased cells such as cancer cells may be different in that DNMT1 alone is not responsible for maintaining abnormal gene hypermethylation and both DNMT1 and DNMT3B may cooperate with this function.


Fig. 1. Schematic procedure for the EpiQuik™ DNMT Activity/Inhibition Assay Ultra Kit (Fluorometric).
Product Details


Fig. 2. EpiQuik DNMT activity and inhibition fluorometric assay. High sensitivity and specificity achieved by using recombinant DNMT1 with the EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Fluorometric).

Principle & Procedure
In the non-radioactive assay with this kit, a universal DNMT substrate is stably coated onto the wells. DNMT enzymes transfer methyl groups to cytosines from Adomet to methylate the DNA substrate. The methylated DNA can be recognized with an anti-5-methylcytosine antibody. The ratio or amount of methylated DNA, which is proportional to the enzyme activity, can then be fluorometrically measured by reading the fluorescence intesntiy in a fluorescence microplate spectrophotometer at excitation 530 +/-20 nm and emission 580 +/-20 nm. The activity of DNMT enzymes is proportional to the RFU (relative fluorescence units) measured.

Easy, Fast, & Flexible
The entire fluorometric assay has easy-to-follow steps for convenience and speed, which can be completed in 3 hours and 45 minutes. The stripwell microplate format allows for a flexible assay in manual or high throughput analysis. 

Safe & Convenient
All the needed reagents, including negative controls and positive controls, for measurement of DNMT activity/inhibition are conveniently packaged in the kit. The direct fluorometric quantification of DNA samples replaces obsolete or inferior methods and eliminates the need for radioactivity, extraction, or chromatography. 

Highly Sensitive & Specific
The novel procedure and proprietary kit compositions allow for an accurate measurement of DNMT activity/inhibition to be achieved with high sensitivity and specificity. The detection limit of the input DNA can be as low as 0.2 µg of nuclear extract or 0.2 ng of purified enzymes, which is ten times better than the predecessor kit.

Product Components

MK1 (10X Wash Buffer)
MK2 (DNMT Assay Buffer)
MK3 (Adomet, 50X)*
MK4 (DNMT Enzyme Control, 50 µg/ml)*
MK5 (Capture Antibody, 1000 µg/ml)*
MK6 (Detection Antibody, 400 µg/ml)*
MK7 (Enhancer Solution)*
MK8 (Fluoro Developer)
MK9 (Fluoro Enhancer)
MK10 (Fluoro Diluter)
8-Well Assay Strips (With Frame)
Adhesive Covering Film
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.

Product Citations

Nugent BM et. al. (May 2015). Brain feminization requires active repression of masculinization via DNA methylation. Nat Neurosci. 18(5):690-7.

Wang B et. al. (January 2015). Exposure to low dose cadmium enhances FL83B cells proliferation through down-regulation of caspase-8 by DNA hypermethylation Toxicol. Res.. (Advance Article) Abstract

Olsvik PA et. al. (July 2014). Impact of dietary selenium on methylmercury toxicity in juvenile Atlantic cod: A transcriptional survey. Chemosphere. 120C:199-205.

Dunn J et. al. (May 2014). Flow-dependent epigenetic DNA methylation regulates endothelial gene expression and atherosclerosis. J Clin Invest.

Zhang D et. al. (October 2012). Homocysteine activates vascular smooth muscle cells by DNA demethylation of platelet-derived growth factor in endothelial cells. J Mol Cell Cardiol. 53(4):487-96.

Epigentek Product Reviews
Related Products

Researchers Also Purchased
EpiQuik Nuclear Extraction Kit
EpiQuik Nuclear Extraction Kit
MethylFlash Methylated DNA Quantification Kit (Fluorometric)
MethylFlash Methylated DNA Quantification Kit (Fluorometric)
EpiQuik Dnmt1 Assay Kit
EpiQuik Dnmt1 Assay Kit
MethylFlash Methylated DNA 5-mC Quantification Kit (Colorimetric)
MethylFlash Methylated DNA 5-mC Quantification Kit (Colorimetric)
Epigentek is an epigenetics company that specializes in epigenetic kits, antibodies, reagents, and services for epigenetic research in DNA methylation, histone modification, and chromatin studies. Terms & Conditions | Privacy Policy | Site Map
Copyright © 2017 Epigentek Group Inc. All rights reserved.