Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 3 hr 45 min
Product nameDNMT Activity Assay Kit (Colorimetric)
See all DNMT kits
Assay typeEnzyme activity (quantitative)
Assay time3h 45m
Species reactivityReacts with: Mouse, Rat, Human, Drosophila melanogaster, Plants
Predicted to work with: Mammals, Fungi
DNMT Activity Quantification Kit (Colorimetric) ab113467 allows the user to colorimetrically measure DNA methyltransferase activity or inhibition at extremely fast speed on a 96-strip well 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.
In the DNMT assay protocol, DNMT enzymes in a sample transfer methyl groups from Adomet to a DNMT DNA substrate coated onto microplate wells. The methylated DNA is bound with a 5-methylcytosine antibody. A secondary antibody binds the 5-methylcytosine antibody, and the enzyme-conjugated secondary antibody is detected using a chromogenic enzyme substrate.
DNMT assay protocol summary:
- add samples, positive controls, and samples with inhibitor to wells
- incubate for 90-120 min, and wash wells three times with wash buffer
- add capture antibody, incubate for 60 min, and wash wells
- add detection antibody, incubate for 30 min, and wash wells
- add enhancer solution, incubate for 30 min, and wash wells
- add developer solution, incubate for 1-10 min
- add stop solution and analyze with a microplate reader
DNMT (DNA methylases) is a family of enzymes responsible for the addition of methyl groups on the cytosine ring at the 5' position of a CpG dinucleotide. The DNMT family is required for the establishment and maintenance of DNA methylation patterns.
Storage instructionsPlease refer to protocols.
Components 48 tests 96 tests 10X Wash Buffer 1 x 14ml 1 x 28ml 8-Well Assay Strips (with Frame) 6 units 12 units Adhesive Covering Film 1 unit 1 unit Adomet, 50X 1 x 60µl 1 x 120µl Capture Antibody, 1000 µg/mL 1 x 5µl 1 x 10µl Detection Antibody, 400 µg/mL 1 x 6µl 1 x 12µl Developer Solution 1 x 5ml 1 x 10ml DNMT Assay Buffer 1 x 4ml 1 x 8ml DNMT Enzyme Control, 50 µg/mL 1 x 6µl 1 x 12µl Enhancer Solution 1 x 6µl 1 x 12µl Stop Solution 1 x 5ml 1 x 10ml
Demonstration of high sensitivity and specificity of the DNMT activity/inhibition assay achieved by using recombinant DNMT1 with ab113467.
Demonstration of high sensitivity and specificity of the DNMT activity assay achieved by using nuclear extracts with ab113467. Nuclear extracts were prepared from MCF-7 cells using ab113474.
ab113467 has been referenced in 9 publications.
- Sales AJ et al. CBD modulates DNA methylation in the prefrontal cortex and hippocampus of mice exposed to forced swim. Behav Brain Res 388:112627 (2020). PubMed: 32348868
- Romero MA et al. Five months of voluntary wheel running downregulates skeletal muscle LINE-1 gene expression in rats. Am J Physiol Cell Physiol 317:C1313-C1323 (2019). PubMed: 31618076
- Mumford PW et al. Skeletal muscle LINE-1 retrotransposon activity is upregulated in older versus younger rats. Am J Physiol Regul Integr Comp Physiol 317:R397-R406 (2019). PubMed: 31188650
- Wang J et al. LMP2A induces DNA methylation and expression repression of AQP3 in EBV-associated gastric carcinoma. Virology 534:87-95 (2019). PubMed: 31220652
- O'Leary VB et al. Long non-coding RNA PARTICLE bridges histone and DNA methylation. Sci Rep 7:1790 (2017). Functional Studies ; Human . PubMed: 28496150
- Panikar CS et al. Presence of DNA methyltransferase activity and CpC methylation in Drosophila melanogaster. Mol Biol Rep N/A:N/A (2015). Functional Studies ; Drosophila melanogaster . PubMed: 26547851
- Albrengues J et al. Epigenetic switch drives the conversion of fibroblasts into proinvasive cancer-associated fibroblasts. Nat Commun 6:10204 (2015). Functional Studies . PubMed: 26667266
- Keller M et al. Global DNA methylation levels in human adipose tissue are related to fat distribution and glucose homeostasis. Diabetologia 57:2374-83 (2014). Functional Studies . PubMed: 25145546
- Gracia A et al. Fatty acid synthase methylation levels in adipose tissue: effects of an obesogenic diet and phenol compounds. Genes Nutr 9:411 (2014). Functional Studies ; Rat . PubMed: 24903834