尿液样本 m6A 定量检测试剂盒（比色法）                                                     货号：A-P-9015

MethylFlash Urine N6-methyladenosine (m6A) Quantification Kit (Colorimetric)

背景资料：Nucleobase m6A, a modified form of adenosine converted by adenosine methyltransferases is widespread in different cellular RNAs and also found in DNA. The biological importance of DNA/RNA m6A-methylation as a major epigenetic modification in phenotype and gene expression has been recognized widely. m6A plays crucial roles in regulating DNA replication, DNA damage, RNA splicing, transposition, transcription, and cellular defense. In humans, the m6A modification is probably catalyzed by a methyltransferase complex METTL3/METTL14 and removed by the α-ketoglutarate (α-KG)- and Fe2+-dependent dioxygenases such as FTO, 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. Urinary excretion of m6A is an indication of the whole body turnover or the degradation of DNA/RNA, especially tRNA. The urinary m6A level can be changed with a change of the bodies’ turnover of m6A DNA/RNA or alteration of cellular DNA/RNA m6A status. A number of studies have indicated that m6A excreted in urine has the potential to act as a cancer biomarker. For example, an elevated level of urinary m6A was observed in colorectal cancer patients with active disease states。

产品描述：In this ELISA-like inhibitory competitive immunoassay, urine samples and the m6A standard are first incubated with a m6A antibody solution and then transferred to strip wells coated with m6A polynucleotide. The well is washed to remove any unbound reagents after incubation and then a detection antibody is added to generate a signal that can be measured colorimetrically by reading the absorbance in a microplate spectrophotometer. Because m6A in the urine sample inhibits the binding of m6A antibody to m6A coated on the well, higher concentrations of m6A in the urine sample lead to a reduced binding of the antibody to the m6A on the well. Therefore the signal or OD intensity measured from the well will be inversely proportional to the amount of m6A in the urine sample and the amount of m6A in the urine sample can be quantified by a comparison with a predetermined m6A standard。

产品特点：

1·Innovative colorimetric assay with easy-to-follow steps for convenience and speed. The entire procedure can be finished within 4 hours；

2·96 strip-well microplate format makes the assay flexible: manual or high throughput analysis；

3.Innovative kit composition enables background signals to be extremely low, which eliminates the need for plate blocking and allows the assay to be simple, accurate, reliable, and consistent；

4.The level of m6A measured in human urine samples using this kit is comparable to that detected by HPLC method；

5.A novel assay principle allows high sensitivity to be achieved. The detection limit can be as low as 0.01 ng/assay well；

6.Low input range of urine for each assay with a volume of 1 to 20 µl and an optimal volume of 5 µl；

7.Optimized antibody and enhancer solutions allow for high specificity to m6A, without cross-reactivity to unmethylated adenosine；

8.Negative control and positive standard are included, which are suitable for quantification of m6A in free form and m6A contained in DNA/RNA fragments from different urine samples。

数据分析： 

操作流程	标准曲线	柱状图分析
尿液样本 m6A 定量检测试剂盒（比色法）流程	将m6A标准品以不同浓度添加到孔中，使用尿液样本 m6A定量检测试剂盒（比色法）来测吸光值	使用尿液样本 m6A定量检测试剂盒（比色法）精确定量来自人类尿液样本不同体积的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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