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Epigenetics |
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| Epigenetics>【Zymo】 |
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Quest 5-hmC™ DNA ELISA Kit(D5425) |
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Quest 5-hmC™ DNA ELISA Kit
About Quest 5-hmC™ DNA ELISA Kit
The 5-methylcytosine (5-mC) modification of DNA in epigenetic regulation has been well studied over the last several decades. However, the role of the so-called “sixth base”, 5-hydroxymethylcytosine (5-hmC), has yet to be defined. Recent studies have associated 5-hmC patterns to transcriptional regulation of genes, which may contribute to normal and disease states of organisms.
The Quest 5-hmC™ DNA ELISA Kit is both sensitive and specific and can be used to accurately detect 5-hmC DNA in a variety of samples. The kit is compatible with a wide range of input DNA including intact vertebrate, plant, and microbial genomic DNA, as well as enzyme-digested and mechanically sheared fragments. The Control DNA Set included with this kit has been calibrated to accurately quantify the percent 5-hmC in sample DNA by use of a standard curve. Also, the fast, streamlined workflow is ideal when analyzing/screening large numbers of samples.
| Equipment |
An incubator and plate reader (w/ 405-450 nm detection) are required. A multi-channel pipettor is recommended. An automated plate washer may be used for blocking and wash steps due to the one-buffer system. |
| Sample Sources |
This ELISA procedure has been optimized for the detection of 5-hmC in purified genomic DNA that is intact, sheared or fragmented in PBS, Tris-EDTA, or water. The product is also compatible with DNA from other sources. |
| Detection |
This system is highly sensitive for 5-hmC DNA and has a lower detection of 0.02% per 100ng input DNA, making it easy to detect even the smallest percentage of 5-hmC in DNA samples. |
Quest 5-hmC™ DNA ELISA Kit is a sandwich-based ELISA format. First, Anti-5-Hydroxymethylcytosine Polyclonal Antibody (anti-5-hmC pAb) is coated to the bottom of a well. Single stranded 5-hmC-containing DNA binds to anti-5-hmC pAb which is then recognized by Anti-DNA HRP Antibody. Addition of HRP Developer will produce a greenish-blue color in the wells containing 5-hmC DNA.
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The Quest 5-hmC™ DNA ELISA Kit can be used to detect 5-hmC in numerous DNA samples with high specificity as evidenced by comparison with LC-MS/MS-MRM analysis. 5-hmC pAb (200 ng/well) was used to quantitate the amount 5-hmC in 100 ng of single-stranded DNA. For this, % 5-hmC was calculated from a standard curve generated using the Control DNA Set. The figure shows a correlation between the % 5-hmC in DNA samples calculated using the Quest 5-hmC™ DNA ELISA Kit (ELISA) and mass spectrometry (MS).
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Sensitive and specific quantitation of 5-hydroxymethylcytosine (5-hmC) DNA from a variety of samples
Ideal for global 5-hmC detection, tissue-specific 5-hmC quantitation, high-throughput compound screening, and more
Streamlined workflow can be completed in as little as 3 hours.
| Quest 5-hmC™ DNA ELISA Kit |
1x96 |
D5425 |
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| Quest 5-hmCDNA ELISA Kit (2x96) |
2x96 |
D5426 |
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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。
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