DNMT Activity Assay Kit (Colorimetric) (ab113467)
製品の概要
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製品名DNMT Activity Assay Kit (Colorimetric)
DNMT キット 製品一覧 -
検出方法Colorimetric
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アッセイタイプEnzyme activity (quantitative)
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全工程の試験時間3h 45m
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種交差性交差種: Mouse, Rat, Human, Drosophila melanogaster, Plants
交差が予測される動物種: Mammals, Fungi -
製品の概要
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 -
特記事項
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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試験プラットフォームMicroplate reader
製品の特性
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保存方法Please refer to protocols.
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内容 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 -
研究分野
画像
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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.
プロトコール
参考文献
This product has been referenced in:
- O'Leary VB et al. Long non-coding RNA PARTICLE bridges histone and DNA methylation. Sci Rep 7:1790 (2017). Functional Studies ; Human . Read more (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 . Read more (PubMed: 26547851) »