DNMT Activity Assay Kit (Colorimetric) (ab113467)
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.
Datasheets and documents
ab113467 has been referenced in 14 publications.
- Aigner GP et al. DNA Methylation and Detoxification in the Earthworm Lumbricus terrestris Exposed to Cadmium and the DNA Demethylation Agent 5-aza-2'-deoxycytidine. Toxics 10:N/A (2022). PubMed: 35202286
- Tarazón E et al. DNMT3B System Dysregulation Contributes to the Hypomethylated State in Ischaemic Human Hearts. Biomedicines 10:N/A (2022). PubMed: 35453616
- Proto MC et al. Modified Adenosines Sensitize Glioblastoma Cells to Temozolomide by Affecting DNA Methyltransferases. Front Pharmacol 13:815646 (2022). PubMed: 35559231
- Anier K et al. Psychostimulant-induced aberrant DNA methylation in an in vitro model of human peripheral blood mononuclear cells. Clin Epigenetics 14:89 (2022). PubMed: 35842682
- Wang W et al. Resveratrol ameliorates toxic effects of cadmium on placental development in mouse placenta and human trophoblast cells. Birth Defects Res 113:1470-1483 (2021). PubMed: 34668346