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
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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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Functional Studies - DNMT Activity Quantification Kit (Colorimetric) (ab113467)Demonstration of high sensitivity and specificity of the DNMT activity/inhibition assay achieved by using recombinant DNMT1 with ab113467.
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Functional Studies - DNMT Activity Quantification Kit (Colorimetric) (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.
