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
Overview
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Product name
DNMT Activity Assay Kit (Colorimetric)
See all DNMT kits -
Detection method
Colorimetric -
Assay type
Enzyme activity (quantitative) -
Assay time
3h 45m -
Species reactivity
Reacts with: Mouse, Rat, Human, Drosophila melanogaster, Plants
Predicted to work with: Mammals, Fungi -
Product overview
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 -
Notes
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.
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Platform
Microplate reader
Properties
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Storage instructions
Please 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 -
Research areas
Images
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Demonstration of high sensitivity and specificity of the DNMT activity/inhibition assay achieved by using recombinant DNMT1 with ab113467.
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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
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SDS download
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Datasheet download
References (9)
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