m6A RNA甲基化定量检测试剂盒(比色法) 货号：A-P-9005
EpiQuik m6A RNA Methylation Quantification Kit (Colorimetric)
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背景资料：N6-methyl-adenosine (m6A) is the most common and abundant modification in RNA molecules present in eukaryotes. The m6A modification is catalyzed by a methyltransferase complex METTL3 and removed by the recently discovered m6A RNA demethylases FTO and ALKBH5, which catalyze m6A demethylation in an α-ketoglutarate (α-KG)- and Fe2+-dependent manner. It was shown that METTL3, FTO, and ALKBH5 play important roles in many biological processes, ranging from development and metabolism to fertility. m6A accounts for more than 80% of all RNA base methylations and exists in various species. m6A is mainly distributed in mRNA and also occurs in non-coding RNA such as tRNA, rRNA, and snRNA. The relative abundance of m6A in mRNA transcripts has been shown to affect RNA metabolism processes such as splicing, nuclear export, translation ability and stability, and RNA transcription. Abnormal m6A methylation levels induced by defects in m6A RNA methylase and demethylase could lead to dysfunction of RNA and diseases. For example, abnormally low levels of m6A in target mRNAs due to increased FTO activity in patients with FTO mutations, through an as-yet undefined pathway, contributes to the onset of obesity and related diseases. The dynamic and reversible chemical m6A modification in RNA may also serve as a novel epigenetic marker of profound biological significance. Therefore, more useful information for a better understanding of m6A RNA methylation levels and distribution on RNA transcripts could benefit diagnostics and therapeutics of disease.
产品描述：In this assay, total RNA is bound to strip wells using a RNA high binding solution. m6A is detected using specific capture and detection antibodies. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The amount of m6A is proportional to the OD intensity measured. Both negative and positive RNA controls are provided in this kit. A standard curve can be performed (range: 0.02 to 1 ng of m6A) or a single quantity of m6A can be used as a positive control. Because m6A content can vary from tissue to tissue, and from normal and diseased states, or vary under treated and untreated conditions, it is advised to run replicate samples to ensure that the signal generated is validated. This kit will allow the user to quantify an absolute amount of m6A and determine the relative m6A RNA methylation states of two different RNA samples。
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| m6A RNA甲基化定量检测试剂盒(比色法)步骤示意图 |
m6A RNA甲基化定量检测试剂盒(比色法)标准曲线
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产品特点：
1·类似于Elisa步骤的比色法检测方法方便快速。整个操作步骤可在3小时45分钟内完成 ；
2·高灵敏，检测的最低下限低至10pg 的 m6A ；
3·独特的结合液容许>70 ngt 的RNA紧紧的结合在孔板上，可以定量检测来自mRNA和ncRNA，如：tRNA，rRNA和snRNA；
4.优化的抗体和增强剂溶液可以特异的定量检测m6A，对于特定片段腺苷浓度范围的未甲基化样本RNA不会产生交叉反应；
5.通用的阴性和阳性对照，对于来自任何种属的m6A都可以实现定量检测；
6.96可拆卸微孔板设计，使得研究人员可根据自己需要选择手动或是高通量模式分析；
7.操作简便，可信，分析条件始终如一。
参考文献：
1.Yuen KC et. al. (January 2016). NIPBL Controls RNA Biogenesis to Prevent Activation of the Stress Kinase PKR. Cell Rep. 14(1):93-102.
2.Zhang C et. al. (April 2016). Hypoxia induces the breast cancer stem cell phenotype by HIF-dependent and ALKBH5-mediated m6A-demethylation of NANOG mRNA. Proc Natl Acad Sci U S A. 113(14):E2047-56.
3.Yin H et. al. (October 2016). Electrochemical immunosensor for N6-methyladenosine detection in human cell lines based on biotin-streptavidin system and silver-SiO2 signal amplification. Biosens Bioelectron.
4.Ishihara Y et. al. (November 2016). Hsa-miR-520d-5p promotes survival in human dermal fibroblasts exposed to a lethal dose of UV irradiation npj Aging and Mechanisms of Disease. (2)
5.Lewinska A et. al. (February 2017). Downregulation of Methyltransferase Dnmt2 Results in Condition-dependent Telomere Shortening and Senescence or Apoptosis in Mouse Fibroblasts.
6.J Cell Physiol.Lewinska A et. al. (August 2017). Reduced levels of methyltransferase DNMT2 sensitize human fibroblasts to oxidative stress and DNA damage that is accompanied by changes in proliferation-related miRNA expression. Redox Biol. 14:20-34.
8.Anna Lewinska et. al. (August 2017). Sulforaphane-Induced Cell Cycle Arrest and Senescence are accompanied by DNA Hypomethylation and Changes in microRNA Profile in Breast Cancer Cells Theranostics. 7(14)
9.Lewinska A et. al. (August 2017). Sulforaphane-Induced Cell Cycle Arrest and Senescence are accompanied by DNA Hypomethylation and Changes in microRNA Profile in Breast Cancer Cells. Theranostics. 7(14):3461-3477.
10.Haiyan Wang et. al. (July 2017). A sensitive photoelectrochemical immunoassay of N6 - methyladenosine based on dual-signal amplification strategy: Ru doped in SiO2 nanosphere and carboxylated g-C3N4 Biosensors and Bioelectronics.
11.Walters BJ et. al. (June 2017). The Role of The RNA Demethylase FTO (Fat Mass and Obesity-Associated) and mRNA Methylation in Hippocampal Memory Formation. Neuropsychopharmacology. 42(7):1502-1510.
12.Elkashef SM et. al. (May 2017). IDH Mutation, Competitive Inhibition of FTO, and RNA Methylation. Cancer Cell. 31(5):619-620.
13.Slobodin B et. al. (April 2017). Transcription Impacts the Efficiency of mRNA Translation via Co-transcriptional N6-adenosine Methylation. Cell. 169(2):326-337.e12.
14.Zhang S et. al. (March 2017). m6A Demethylase ALKBH5 Maintains Tumorigenicity of Glioblastoma Stem-like Cells by Sustaining FOXM1 Expression and Cell Proliferation Program Cancer Cell.
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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