m6A DNA甲基化定量检测试剂盒（比色法） 货号：A-P-9010
MethyFlash m6A DNA Methylation ELISA Kit (Colorimetric)
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背景资料：N6-methyladenosine (m6A) is the most common and abundant modification on RNA molecules present in eukaryotes. DNA m6A is also identified in multicellular eukaryotes including Caenorhabditis elegans and Drosophila melanogaster, and furthermore identified in higher eukaryotes including plants, mouse and human cells. m6A plays crucial roles in regulating DNA replication, transposition, transcription, and cellular defense. In humans, the DNA m6A modification is most likely catalyzed by a methyltransferase complex METTL3 and removed by the α-ketoglutarate (α-KG)- and Fe2+-dependent dioxygenases such as ALKBH5 and TET-like enzymes. It was shown that METTL3 and α-KG /Fe2+-dependent dioxygenases play important roles in many biological processes, ranging from development and metabolism to fertility. The dynamic and reversible chemical m6A modification on DNA may also serve as a novel epigenetic marker of profound biological significance. Down-regulation of m6A modification was first characterized in human cancer cells and tissues, relative to their normal controls. m6A is found to be the most regulated DNA modification in cancers. In addition to the regulation in cancer cells, relative to the primary cell/tissues which contain quite low amounts of DNA m6A (<0.001%), a hundreds-fold increase of m6A modification was found for in vitro cultured human cells (0.03%-0.22%). Therefore, identifying m6A DNA methylation levels and distribution on DNA could advance understanding of epigenetic regulation of biological process at the genomic level, and further provide useful information for improving diagnostics and therapeutics of disease。
产品描述：This kit contains all reagents necessary for the quantification of m6A in DNA. In this assay, DNA is bound to strip wells using DNA high binding solution. m6A is detected using capture and detection antibodies. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The amount of m6A is proportional to the OD intensity measured。
产品特点：
1·Colorimetric assay with easy-to-follow steps for convenience and speed. The entire procedure can be completed within 3 hours and 45 minutes；
2·High sensitivity, of which the detection limit can be as low as 5 pg of m6A；
3.Unique binding solution allows DNA >100 nts to be tightly bound to the wells, which enables quantification of m6A from both intact DNA or fragmented DNA such as ChIPed DNA；
4.Optimized antibody and enhancer solutions allow high specificity to m6A, with no cross-reactivity to unmethylated adenosine within the indicated concentration range of the sample DNA；
5.Universal positive and negative controls are included, which are suitable for quantifying m6A from any species；
6.Strip-well microplate format makes the assay flexible for manual or high throughput analysis；
7.Simple, reliable, and consistent assay conditions；
8.Flexible - Strip-well microplate format makes the assay available for manual or high throughput analysis。
数据分析：
| 操作流程 |
标准曲线 |
柱状图分析 |
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| m6A DNA甲基化定量检测试剂盒（比色法）操作步骤 |
将m6A标准品以不同浓度添加到孔中，使用m6A DNA甲基化定量检测试剂盒（比色法）来测 |
使用m6A DNA甲基化定量检测试剂盒（比色法）精确定量来自人类不同样本DNA的m6A 水平。结果与MS-LC一致。甚至更好。 |
参考文献：
1.Lewinska A et. al. (February 2017). Downregulation of Methyltransferase Dnmt2 Results in Condition-dependent Telomere Shortening and Senescence or Apoptosis in Mouse Fibroblasts. J Cell Physiol.
保存建议：在接收到艾德寄出的试剂盒后请按照说明书建议，使用不同的保存条件或温度来保存试剂盒内组分。
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5-hmC是近年来在动物组织中发现的，由胞嘧啶修饰而来。5-hmC在表观遗传学上的功能可能与5-甲基化胞嘧啶(5-mC)不同。尽管到现在为止还不确知其功能，有研究者猜测它在调控基因的表达与关闭过程中起着重要的作用。5-mC的发现让我们不得不重新评估DNA甲基信息，也不得不监测人类的健康组织和病理组织之间5-mC相对分布的差异。在EPI公司的MethylFlash技术之前，我们还没有发现任何直接的常规方法来检测5- hmC,以及区分5-hmC和5-mC
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