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EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric)

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For absorbance-based measurement of DNA methyltransferase activity or inhibition

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EpiQuik DNMT activity and inhibition colorimetric assay: (a) High sensitivity and specificity achieved by using recombinant DNMT1 with the EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric); (b) High sensitivity and specificity achieved by using nuclear extract with the EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric). Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit (Cat. No. OP-0002).
Schematic procedure for the EpiQuik™ DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric).
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-3009-4848 assays $327.00 
P-3009-9696 assays $551.00 
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 is a complete set of optimized buffers and reagents that allows the experimenter to colorimetrically 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 five times more sensitive. The kit has the following advantages and features:

  • Colorimetric 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 colorimetric assay without radioactivity, extraction, and chromatography.
  • The ultra-sensitive detection limit can be as low as 0.5 µg of nuclear extract or 0.5 ng of purified enzymes, which is five 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.

Product Details


Fig. 2. EpiQuik DNMT activity and inhibition colorimetric assay: (a) High sensitivity and specificity achieved by using recombinant DNMT1 with the EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric); (b) High sensitivity and specificity achieved by using nuclear extract with the EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric). Nuclear extracts were prepared from MCF-7 cells using the EpiQuik Nuclear Extraction Kit (Cat. No. OP-0002).

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 colorimetrically measured through an ELISA-like reaction by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The activity of DNMT enzymes is proportional to the intensity of optical density measured.

Easy, Fast, & Flexible
The entire colorimetric 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 colorimetric 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 0.5 µg of nuclear extract or 0.5 ng of purified enzymes, which is five times better than the predecessor kit.

Product Components

MU1 (10X Wash Buffer)
MU2 (DNMT Assay Buffer)
MU3 (Adomet, 50X)*
MU4 (DNMT Enzyme Control, 50 µg/ml)*
MU5 (Capture Antibody, 1000 µg/ml)*
MU6 (Detection Antibody, 400 µg/ml)*
MU7 (Enhancer Solution)*
MU8 (Developer Solution)
MU9 (Stop Solution)
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

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Fan Yang et. al. (December 2016). Casticin inhibits the activity of transcription factor Sp1 and the methylation of RECK in MGC803 gastric cancer cells Experimental and Therapeutic Medicine. 13(2):745-750.

Sanchez OF et. al. (December 2016). Lead (Pb) exposure reduces global DNA methylation level by non-competitive inhibition and alteration of dnmt expression. Metallomics.

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Herencia C et. al. (June 2016). Procaine Inhibits Osteo/Odontogenesis through Wnt/β-Catenin Inactivation. PLoS One. 11(6):e0156788.

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Sun R et. al. (November 2015). Deoxyribonucleic acid (DNA) methyltransferase contributes to p16 promoter CpG island methylation in lung adenocarcinoma with smoking Int J Clin Exp Med. 8(9):15773-15779.

Naselli F et. al. (October 2015). Phytochemical Indicaxanthin Inhibits Colon Cancer Cell Growth and Affects the DNA Methylation Status by Influencing Epigenetically Modifying Enzyme Expression and Activity. J Nutrigenet Nutrigenomics. 8(3):114-27.

Pushpakumar S et. al. (July 2015). DNA hypermethylation in hyperhomocysteinemia contributes to abnormal extracellular matrix metabolism in the kidney. FASEB J.

Li JJ et. al. (June 2015). Homocysteine Triggers Inflammatory Responses in Macrophages through Inhibiting CSE-H2S Signaling via DNA Hypermethylation of CSE Promoter. Int J Mol Sci. 16(6):12560-12577.

Pathania R et. al. (April 2015). DNMT1 is essential for mammary and cancer stem cell maintenance and tumorigenesis. Nat Commun. 6:6910.

Zeng F et. al. (March 2015). Dynamic change of DNA methylation and cell redox state at different micropropagation phases in birch. Trees. :1-14.

Ishikawa K et. al. (February 2015). Long-term pancreatic beta cell exposure to high levels of glucose but not palmitate induces DNA methylation within the insulin gene promoter and represses transcriptional activity. PLoS One. 10(2):e0115350.

Bian Y et. al. (February 2015). Epigenetic Regulation of miR-29s Affects the Lactation Activity of Dairy Cow Mammary Epithelial Cells. J Cell Physiol.

Cho SH et. al. (January 2015). SIRT1 deficiency in microglia contributes to cognitive decline in aging and neurodegeneration via epigenetic regulation of IL-1β. J Neurosci. 35(2):807-18.

Cheng C et. al. (December 2014). 'Default' generated neonatal regulatory T cells are hypomethylated at conserved non-coding sequence 2 and promote long-term cardiac allograft survival. Immunology. 143(4):618-30.

Zhang S et. al. (December 2014). The Effect of Hypoxia Preconditioning on DNA Methyltransferase and PP1γ in Hippocampus of Hypoxia Preconditioned Mice. High Alt Med Biol. 15(4):483-90.

Cardenas H et. al. (November 2014). TGF-β induces global changes in DNA methylation during the epithelial-to-mesenchymal transition in ovarian cancer cells. Epigenetics.

Chen S et. al. (November 2014). Identifying Novel Selective Non-Nucleoside DNA Methyltransferase 1 Inhibitors through Docking-Based Virtual Screening. J Med Chem. 57(21):9028-41.

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Customer Reviews

Rating by 2********@qq.com Reviewed on: Tuesday 22 August, 2017
Application Description
This product is for determine the DNMT activity, the price is great, with this perfect product
Rating by L**@uab.edu Verified Purchase Reviewed on: Monday 19 December, 2016
Application Description
We used EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (p-3009) and EpiQuik HDAC Activity/Inhibition Assay Kit (p-4002) to check DNMT and HDAC activity in tumor cells.

Pros: They are very easy to work with. Protocols are clearly delineated the detailed procedures.
Cons: Too much steps and solutions, which may cause confusion and mistakes.

Other Thoughts
1) DNMT kit should have positive control included.
2) There is a concern for how to determine internal HDAC and DNMT quantity (protein level) to normalize all samples.

How friendly was the user guide? (1 – worst, 5 – best)
4

How professional was the appearance and presentation of the product? (1 – worst, 5 – best)
5

How would you rate the overall product? (1 – worst, 5 – best)
4.5
Epigentek Product Reviews
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