總體RNA甲基化極易定量檢測試劑盒(熒光法) 貨號:A-P-9009 MethylFlash 5-mC RNA Methylation ELISA Easy Kit (Fluorometric) 背景資料: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。 產品描述: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。 產品特點: 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。 數據分析: 操作流程 | 標準曲線 | 柱狀圖分析 |  |  |  | 總體RNA甲基化極易定量檢測試劑盒(熒光法)步驟 | 使用5-mC陽性對照繪制標準曲線 | 使用總體RNA甲基化極易定量檢測試劑盒(熒光法)精確定量來自各種種屬的RNA。結果與MS-LC一致。甚至更好。 |
保存建議:在接收到艾德寄出的試劑盒后請按照說明書建議,使用不同的保存條件或溫度來保存試劑盒內組分。 定購信息: 產品名稱 | 規格 | 操作手冊 | 貨號 | 詢價 | 總體RNA甲基化極易定量檢測試劑盒(熒光法) | 96次 | 英文PDF-中文PDF | A-P-9009-96 |  | 總體RNA甲基化極易定量檢測試劑盒(熒光法) | 48次 | 英文PDF-中文PDF | A-P-9009-48 | | 總組蛋白H3K27乙酰化定量檢測試劑盒(比色法) | 48次 | PDF | A-P-4059-48 | | 總組蛋白H3K27乙酰化定量檢測試劑盒(比色法) | 96次 | PDF | A-P-4059-96 | | EpiNext 高靈敏重亞硫酸鹽測序試劑盒(Illumina) | 12次 | PDF | A-P-1056A-12 | EpiNext 高靈敏重亞硫酸鹽測序試劑盒(Illumina) | 24次 | PDF | A-P-1056A-24 |  | 96孔DNA甲基化極速修飾試劑盒(磁珠法) | 96次 | 英文PDF-中文PDF | A-P-1050-096 | | 96孔DNA甲基化極速修飾試劑盒(磁珠法) | 192次 | 英文PDF-中文PDF | A-P-1050-192 | | 抗人C5a/C5a desArg單克隆抗體(克隆號2942) | 100ug | | C-HM2078 | | 抗人TNF-R II單克隆抗體(克隆號80M2),FITC標記 | 100ug | | C-HM2022F | | 抗人TNF-R II單克隆抗體(克隆號MR2-1),生物素標記 | 50ug | | C-HM2008 | | 抗人TNF-R I單克隆抗體(克隆號MR1-2),生物素標記 | 50ug | | C-HM2006 | | 抗小鼠TREM-2單克隆抗體(克隆號6E9) | 100ug | PDF | C-HM1129F |  | m6A RNA甲基化定量檢測試劑盒(比色法) | 96 次 | 英文PDF--中文PDF | A-P-9005-96 | | RNA甲基化修飾試劑盒 | 50 次 | 英文PDF--中文PDF | A-P-9003-050 | | EZ RNA甲基化修飾試劑盒 | 50 次 | 英文PDF--中文PDF | A-R5001 | |
表觀遺傳學產品全面解決方案列下: 推薦閱讀: 關于5-hmC 5-hmC是近年來在動物組織中發現的,由胞嘧啶修飾而來。5-hmC在表觀遺傳學上的功能可能與5-甲基化胞嘧啶(5-mC)不同。盡管到現在為止還不確知其功能,有研究者猜測它在調控基因的表達與關閉過程中起著重要的作用。5-mC的發現讓我們不得不重新評估DNA甲基信息,也不得不監測人類的健康組織和病理組織之間5-mC相對分布的差異。在EPI公司的MethylFlash技術之前,我們還沒有發現任何直接的常規方法來檢測5- hmC,以及區分5-hmC和5-mC 5-hmC 和 5-mC的區別 時下常用的DNA甲基化分析方法包括限制內切酶酶切和亞硫酸氫鹽或MeDIP介導的MS-PCR和測序,這些技術都不適合用來檢測5-hmC,因為它與5-mC事實上很難用這類方法區分開來。為了解決這個問題,EPIk研制了MethylFlash羥甲基化DNA定量試劑盒(熒光法)。本試劑盒提供了一種很經濟的方法來檢測5-羥甲基化胞嘧啶,并且區分5-hmC, 5-mC, 和 C,使得研究者能夠重新評估他們的DNA甲基化信息,也能夠在新樣品中尋找DNA羥甲基化。 |