The EpiQuik™ DNA Methyltransferase Activity/Inhibition Assay Kit is a convenient package of tools that allows the experimenter to measure Dnmt activity or inhibition at tremendously fast speeds and consistency, superior and safer than all other current methods. 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 (Dnmt) Assay Kit is suitable for measuring Dnmt activity/inhibition from a broad range of species including mammalian cells/tissues, plants, and bacteria.
WHY CHOOSE THE EPIQUIK™ DNA METHYLTRANSFERASE ACTIVITY/INHIBITION ASSAY KIT?
- 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 methyl group to cytosine from Adomet to methylate DNA substrate. The methylated DNA can be recognized with 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.
Analysis of Dnmt activity in Hela nuclear extract.
M1 (10x wash buffer)
M2 (Dnmt assay buffer)
M3 (Adomet, 8 mM)*
M4 (Dnmt positive control)*
M5 (capture antibody)*
M6 (detection antibody 200 µg/ml)*
M7 (developing solution)
M8 (stop solution)
8 well substrate-coated strip (with frame)
* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.
User Guide & MSDS
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing firstname.lastname@example.org along with your contact information and institution name.
References & Citations
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. PubMed Abstract
Liao, Yi-Jen et al. (Sep 9, 2008). Characterization of a glycine N-methyltransferase gene knockout mouse model for hepatocellular carcinoma: Implications of the gender disparity in liver cancer susceptibility. International Journal of Cancer Epub ahead of print. Abstract
Guastafierro, Tiziana et al. (Aug 8, 2008). CCCTC-binding Factor Activates PARP-1 Affecting DNA Methylation Machinery. J Biol Chem. 283(32): 21873-21880. PubMed Abstract
Gravina, Giovanni Luca. et al. (May 15, 2008). Chronic azacitidine treatment results in differentiating effects, sensitizes against bicalutamide in androgen-independent prostate cancer cells. The Prostate 68(7): 793-801. PubMed Abstract
Martínez-Chantar, M. Luz et al. (April 2008). Loss of the Glycine N-Methyltransferase Gene Leads to Steatosis and Hepatocellular Carcinoma in Mice. The Prostate 47(4): 1191-1199. PubMed Article
Suzuki, Maiko et al. (Mar 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, Chi et al. (Feb 2008). Plasma S-adenosylhomocysteine is a better biomarker of atherosclerosis than homocysteine in apolipoprotein E-deficient mice fed high dietary methionine. The Journal of Nutrition 138(2): 311-5. PubMed Abstract
Roll, J Devon et al. (Jan 2008). DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines. Mol Cancer 7: 15. Full PDF Article
Jagadeesh, Shankar et al. (Oct 2007). Mahanine reverses an epigenetically silenced tumor suppressor gene RASSF1A in human prostate cancer cells. Biochemical and Biophysical Research Communications 362(1): 212-7. PubMed Abstract
Miyazaki, Tamana et al. (Oct 2007). E-cadherin gene promoter hypermethylation in H. pylori-induced enlarged fold gastritis. Helicobacter 12(5): 523-531. PubMed Abstract
Kodach, Liudmila L. et al. (2007). Lovastatin induces the demethylation of the Bone Morphogenetic Protein 2 promoter in colorectal cancer cell lines. The Role of the Bone Morphogenetic Protein Pathway in Colorectal Cancer 2007: 115-28. Full PDF Article
Paluszczak, Jarosław et al. (May 2007). Modulation of the activity of DNA methyltransferases by plant phenolic compounds. Zjazd Polskiego Towarzystwa Biochemicznego Sympozjum E: P5.7. Science24 Abstract