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EpiQuik DNA Methyltransferase (DNMT) Activity/Inhibition Assay Kit


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Schematic procedure for using the EpiQuik DNA Methyltransferase (DNMT) Activity/Inhibition Assay Kit.
Histone extracts were prepared from MDA-231 cells using the EpiQuik Total Histone Extraction Kit (Cat. No. OP-0006) and the amount of dimethyl-H3-K4 was measured using the EpiQuik Dimethyl Histone H3-K4 Quantification kit (Colorimetric).
HSC-3 cells were incubated with/without zebularine (220 mM) for 48 hours. Nuclear proteins were extracted and total DNMT activity was measured.
Input Type: DNA
Research Area: DNA Methylation
Target Application: Activity Measurement
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-3001-148 assays $339.00 
P-3001-296 assays $569.00 
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Product Overview

We recommend using a newer version of this product: EpiQuik DNMT Activity/Inhibition ELISA Easy Kit (Colorimetric)

The EpiQuik™ DNA Methyltransferase Activity/Inhibition Assay Kit is a complete set of optimized buffers and reagents that allows the experimenter to measure DNA methyltransferase activity or inhibition at extremely fast speeds. 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™ DNA Methyltransferase Activity/Inhibition Assay Kit is suitable for measuring DNMT activity/inhibition from a broad range of species including mammalian cells/tissues, plants, and bacteria. 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.
  • Strip microplate format makes the assay flexible: manual or high throughput analysis.
  • Extremely simple, reliable, and consistent assay conditions.

Principle & Procedure
The EpiQuik™ DNA Methyltransferase Activity/Inhibition Assay Kit is designed for measuring total DNMT activity (de novo, maintenance). In an assay with this kit, the unique cytosine-rich DNA substrate is stably coated on the strip wells. These wells are specifically treated to have a high DNA absorption ability. DNMT enzymes transfer a methyl group to cytosine from Adomet to methylate 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 enzyme activity, can then be colorimetrically quantified through an ELISA-like reaction.

Frequently Asked Q's

1. Can protein extracts from frozen tissue stored at -80°C be used with this kit?
No, protein extracts should be used from fresh tissue.

2. What is the minimum amount of Dnmt protein required for a successful assay?
The minimum amount of DNMT protein is about 0.2 µg for a successful assay.

3. Do I have to do all the assays at once with the kit or can I do them in portions?
Yes, you can do them in portions by using only the reagent amounts you need, including diluting M5 and M6 as needed.

4. How do you aspirate the washing solution from the wells?
By using pipette tips.

5. How long can M5 and M6 be stored for after dilution?
1 or 2 days.

6. Can this kit distinguish Dnmt1 and Dnmt3a/3b?
The P-3001 kit cannot distinguish between Dnmt1 and Dnmt3a/3b. It will work well with both enzymes.

7. How are the wells coated?
The wells are coated with a proprietary Dnmt binding substance (high affinity).

8. Is there any reason why I cannot start with recombinant DNMT1, incubate with my inhibitors, and then use this to proceed with the assay?
This kit is suitable for measuring Dnmt activity of both cell extract and recombinant Dnmts (including Dnmt1) in the absence or presence of inhibitors. The capture mAb included in this kit is against converted (methylated) substrate.

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 along with your contact information and institution name.

[Safety Data Sheet]
Product Citations

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Montes de Oca A et. al. (September 2010). High-phosphate-induced calcification is related to SM22α promoter methylation in vascular smooth muscle cells. J Bone Miner Res. 25(9):1996-2005.

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Xiang H et. al. (May 2010). Single base-resolution methylome of the silkworm reveals a sparse epigenomic map. Nat Biotechnol. 28(5):516-20.

Li Y et. al. (May 2010). Glucose restriction can extend normal cell lifespan and impair precancerous cell growth through epigenetic control of hTERT and p16 expression. FASEB J. 24(5):1442-53.

Kastl L et. al. (April 2010). Effects of decitabine on the expression of selected endogenous control genes in human breast cancer cells. Mol Cell Probes. 24(2):87-92.

Sheikh KD et. al. (March 2010). Fluorescent epigenetic small molecule induces expression of the tumor suppressor ras-association domain family 1A and inhibits human prostate xenograft. J Med Chem. 53(6):2376-82.

Paluszczak J et. al. (February 2010). The effect of dietary polyphenols on the epigenetic regulation of gene expression in MCF7 breast cancer cells. Toxicol Lett. 192(2):119-25.

Majid S et. al. (January 2010). Genistein reverses hypermethylation and induces active histone modifications in tumor suppressor gene B-Cell translocation gene 3 in prostate cancer. Cancer. 116(1):66-76.

Duthie SJ et. al. (January 2010). Folate deficiency alters hepatic and colon MGMT and OGG-1 DNA repair protein expression in rats but has no effect on genome-wide DNA methylation. Cancer Prev Res (Phila). 3(1):92-100.

Watson JA et. al. (October 2009). Generation of an epigenetic signature by chronic hypoxia in prostate cells. Hum Mol Genet. 18(19):3594-604.

Ding L et. al. (June 2009). Camptothecin-induced cell proliferation inhibition and apoptosis enhanced by DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine. Biol Pharm Bull. 32(6):1105-8.

L. Ding et. al. (June 2009). Camptothecin-induced cell proliferation inhibition and apoptosis enhanced by DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine. Biol Pharm Bull.. 32(6):1105-8. Full PDF Article

Gaymes TJ et. al. (May 2009). Inhibitors of poly ADP-ribose polymerase (PARP) induce apoptosis of myeloid leukemic cells: potential for therapy of myeloid leukemia and myelodysplastic syndromes. Haematologica. 94(5):638-46.

Majid S et. al. (April 2009). BTG3 tumor suppressor gene promoter demethylation, histone modification and cell cycle arrest by genistein in renal cancer. Carcinogenesis. 30(4):662-70.

Zampieri M et. al. (March 2009). Parp1 localizes within the Dnmt1 promoter and protects its unmethylated state by its enzymatic activity. PLoS One. 4(3):e4717.

M. Zampieri et. al. (March 2009). Parp1 localizes within the Dnmt1 promoter and protects its unmethylated state by its enzymatic activity. PLoS ONE. 4(3):e4717. Full PDF Article

Liao YJ et. al. (February 2009). Characterization of a glycine N-methyltransferase gene knockout mouse model for hepatocellular carcinoma: Implications of the gender disparity in liver cancer susceptibility. Int J Cancer. 124(4):816-26.

Wang X et. al. (January 2009). RAF may induce cell proliferation through hypermethylation of tumor suppressor gene promoter in gastric epithelial cells. Cancer Sci. 100(1):117-25.

Lim SO et. al. (December 2008). Epigenetic changes induced by reactive oxygen species in hepatocellular carcinoma: methylation of the E-cadherin promoter. Gastroenterology. 135(6):2128-40, 2140.e1-8.

Guastafierro T et. al. (August 2008). CCCTC-binding factor activates PARP-1 affecting DNA methylation machinery. J Biol Chem. 283(32):21873-80.

Suzuki M et. al. (July 2008). Zebularine-induced reduction in VEGF secretion by HIF-1α degradation in oral squamous cell carcinoma. Mol Med Rep. 1(4):465-71.

Gravina GL et. al. (May 2008). Chronic azacitidine treatment results in differentiating effects, sensitizes against bicalutamide in androgen-independent prostate cancer cells. Prostate. 68(7):793-801.

Martínez-Chantar ML et. al. (April 2008). Loss of the glycine N-methyltransferase gene leads to steatosis and hepatocellular carcinoma in mice. Hepatology. 47(4):1191-9.

M. Suzuki et. al. (March 2008). Zebularine-induced reduction in VEGF secretion by HIF-1α degradation in oral squamous cell carcinoma. Molecular Medicine Reports. 1:465-471. Full PDF Article

Liu C et. al. (February 2008). Plasma S-adenosylhomocysteine is a better biomarker of atherosclerosis than homocysteine in apolipoprotein E-deficient mice fed high dietary methionine. J Nutr. 138(2):311-5.

Roll JD et. al. (January 2008). DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines. Mol Cancer. 7:15.

J.D. Roll et. al. (January 2008). DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines. Molecular Cancer. 7(15) Full PDF Article

Jagadeesh S et. al. (October 2007). Mahanine reverses an epigenetically silenced tumor suppressor gene RASSF1A in human prostate cancer cells. Biochem Biophys Res Commun. 362(1):212-7.

Miyazaki T et. al. (October 2007). E-cadherin gene promoter hypermethylation in H. pylori-induced enlarged fold gastritis. Helicobacter. 12(5):523-31.

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