总体RNA甲基化极易定量检测试剂盒(荧光法) 货号:A-P-9009 MethylFlash 5-mC RNA Methylation ELISA Easy Kit (Fluorometric) 背景资料:5-methylcytosine (5-mC) in DNA occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring by DNA methyltransferases. This process has been well studied and is generally associated with repression of gene expression. It was also observed that in humans, 5-mC occurs in various RNA molecules including tRNAs, rRNAs, mRNAs and non-coding RNAs (ncRNAs). 5-mC seems to be enriched in some classes of ncRNA, but relatively depleted in mRNAs. Levels of 5-mC are variable in animal genomes, ranging from undetectable amounts in some insects to about 0.1-0.45% of total RNA in human cells. The majority (83%) of 5-mC sites were found in mRNAs. Within these transcripts 5-mC appears to be depleted within protein coding sequences but enriched in 5’ and 3’ UTRs. Two different methyltransferases, NSUN2 and Dnmt2 are known to catalyze 5-mC modification in eukaryotic RNA. There has been strong evidence that RNA cytosine methylation affects the regulation of various biological processes such as RNA stability and mRNA translation. Furthermore, loss of 5-mC in vault RNAs causes aberrant processing into Argonaute-associated small RNA fragments that can function as microRNAs. Thus, impaired processing of vault ncRNA may contribute to the etiology of human disorders related to NSun2-defciency。 产品描述:This kit contains all reagents necessary for the quantification of global 5-mC RNA methylation. In the assay, RNA is bound to strip-wells that are specifically treated to have a high nucleic acid affinity. 5-mC in RNA is detected using capture and detection antibodies and then quantified fluorometrically by reading the fluorescence in a microplate spectrophotometer. The percentage of 5-mC RNA is proportional to the fluorescence intensity measured。 产品特点: 1·Fast - The entire procedure only needs 2 hours and 40 minutes; 2·Robust - The kit composition allows the assay to have a large “signal window” with less variation between replicates ; 3.Convenient - Inherently low background noise, thereby eliminating the need for plate blocking steps; 4.Sensitive - Detection limit can be as low as 0.02% of 5-mC RNA from 200 ng of input RNA; 5.Specific - High specificity to 5-mC, with no cross-reactivity to unmethylated cytosine or hydroxymethylated cytosine within the indicated concentration range of the sample RNA; 6.Universal - Positive and negative controls allow detection of 5-mC RNA methylation in any species; 7.Accurate - Optimized positive controls that can be fractionalized in percentage scale, allowing the assay to be more accurate and highly comparable with HPLC-MS analysis; 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 | | | | 总体RNA甲基化极易定量检测试剂盒(荧光法)步骤 | 使用5-mC阳性对照绘制标准曲线 | 使用总体RNA甲基化极易定量检测试剂盒(荧光法)精确定量来自各种种属的RNA。结果与MS-LC一致。甚至更好。 |
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。 定购信息: 产品名称 | 规格 | 操作手册 | 货号 | 询价 | 总体RNA甲基化极易定量检测试剂盒(荧光法) | 96次 | 英文PDF-中文PDF | A-P-9009-96 | | 总体RNA甲基化极易定量检测试剂盒(荧光法) | 48次 | 英文PDF-中文PDF | A-P-9009-48 | | 总组蛋白H3K27乙酰化定量检测试剂盒(比色法) | 48次 | PDF | A-P-4059-48 | | 总组蛋白H3K27乙酰化定量检测试剂盒(比色法) | 96次 | PDF | A-P-4059-96 | | EpiNext 高灵敏重亚硫酸盐测序试剂盒(Illumina) | 12次 | PDF | A-P-1056A-12 | EpiNext 高灵敏重亚硫酸盐测序试剂盒(Illumina) | 24次 | PDF | A-P-1056A-24 | | 96孔DNA甲基化极速修饰试剂盒(磁珠法) | 96次 | 英文PDF-中文PDF | A-P-1050-096 | | 96孔DNA甲基化极速修饰试剂盒(磁珠法) | 192次 | 英文PDF-中文PDF | A-P-1050-192 | | 抗人C5a/C5a desArg单克隆抗体(克隆号2942) | 100ug | | C-HM2078 | | 抗人TNF-R II单克隆抗体(克隆号80M2),FITC标记 | 100ug | | C-HM2022F | | 抗人TNF-R II单克隆抗体(克隆号MR2-1),生物素标记 | 50ug | | C-HM2008 | | 抗人TNF-R I单克隆抗体(克隆号MR1-2),生物素标记 | 50ug | | C-HM2006 | | 抗小鼠TREM-2单克隆抗体(克隆号6E9) | 100ug | PDF | C-HM1129F | | m6A RNA甲基化定量检测试剂盒(比色法) | 96 次 | 英文PDF--中文PDF | A-P-9005-96 | | RNA甲基化修饰试剂盒 | 50 次 | 英文PDF--中文PDF | A-P-9003-050 | | EZ RNA甲基化修饰试剂盒 | 50 次 | 英文PDF--中文PDF | A-R5001 | |
表观遗传学产品全面解决方案列下: 推荐阅读: 关于5-hmC 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 5-hmC 和 5-mC的区别 时下常用的DNA甲基化分析方法包括限制内切酶酶切和亚硫酸氢盐或MeDIP介导的MS-PCR和测序,这些技术都不适合用来检测5-hmC,因为它与5-mC事实上很难用这类方法区分开来。为了解决这个问题,EPIk研制了MethylFlash羟甲基化DNA定量试剂盒(荧光法)。本试剂盒提供了一种很经济的方法来检测5-羟甲基化胞嘧啶,并且区分5-hmC, 5-mC, 和 C,使得研究者能够重新评估他们的DNA甲基化信息,也能够在新样品中寻找DNA羟甲基化。 |