DNA羟甲基化免疫共沉淀试剂盒 货号：A-P-1038
EpiQuik Hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit
背景资料：EpiQuik 羟甲基化DNA免疫沉淀(hMeDIP)试剂盒是包含最佳溶液和试剂一套完整实验组件，能成功执行hMeDIP程序，富集DNA片段；还包含5-羟甲基胞嘧啶用于高通量分析来自任何物种DNA。本试剂盒沉淀的羟甲基化DNA常被用于多种下游分析，包括：PCR (hMeDIP-PCR)，微阵列(hMeDIP-chip)来研究细胞或者组织中特异的羟甲基化基因。
产品描述：EpiQuik 羟甲基化DNA免疫沉淀(hMeDIP)试剂盒含有能成功执行hMeDIP所有试剂，利用来自哺乳动物细胞或者组织中的DNA来完成实验。本试剂盒含有阳性对照DNA片断，阴性对照无免疫性的IgG和对照引物，这些可以用来展示羟甲基化DNA的富集功效。阳性对照DNA含有5-hmC，可以通过5-hmC抗体来进行免疫沉淀，但是不能通过无免疫性IgG来实现，在hMeDIP,免疫沉淀过程中，5-hmC富集在微孔板里的DNA片断，在最佳反应条件下进行处理，整个实验完成需要3个小时。免疫沉淀的羟甲基化的DNA然后被清洗，释放和洗提。洗提的DNA能用于多种下游应用，包括：PCR (hMeDIP-PCR)，微阵列(hMeDIP-chip)。
产品特点：
1、快速方便的操作，整个实验（从上样准备好使用羟甲基化DNA开始）不超过3个小时，其中手工操作时间不超过20分钟；
2、微孔板形式让实验更灵活：适用于手工操作或者高通量分析；
3、高富集效率阳性对照/阴性对照> 1000，每次反应最低上样量为0.1ug，高度重复性。兼容于下游实验分析，包括：hMeDIP-PCR and hMeDIP-chip。
实验图片：
| 实验流程 |
数据展示 |
数据展示 |
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| 采用DNA羟甲基化免疫共沉淀试剂盒流程图 |
Sensitive detection of gene-specific hydroxymethylation by hMeDIP-PCR | Human brain DNA (500 ng) was fragmented to 200-600 bps. The fragmented DNA was used for hydroxymethylated DNA enrichment with the EpiQuik™ hMeDIP Kit. Eluted DNA was analyzed by real time PCR with primers specifically for OCT4 or GAPDH sequences in the promoter regions. Results show that the promoter region is hydroxymethylated in OCT4 but not in GAPDH. Fold-enrichment represents the amount of recovered DNA and was calculated based on the Ct Value. |
Selective enrichment of hydroxymethyated DNA with the EpiQuik™ hMeDIP Kit | 50 pg of unmethylated, methylated, and hydroxymethylated DNA control were each spiked into fragmented human genomic DNA (500 ng). hMeDIP was processed with the 5-hmC antibody and non-immune IgG included in the kit. Eluted DNA was analyzed by real time PCR with the control primers included in the kit to detect the presence of spiked control DNA. Fold-enrichment represents the amount of recovered control DNA and was calculated based on the Cts. |
对比数据：
参考文献：Fernandez AF et. al. (June 2018). Loss of 5hmC identifies a new type of aberrant DNA hypermethylation in glioma. Hum Mol Genet.
Duraisamy AJ et. al. (December 2017). Crosstalk Between Histone and DNA Methylation in Regulation of Retinal Matrix Metalloproteinase-9 in Diabetes. Invest Ophthalmol Vis Sci. 58(14):6440-6448.
Rea M et. al. (November 2017). Selective inhibition of CTCF binding by iAs directs TET-mediated reprogramming of 5-hydroxymethylation patterns in iAs-transformed cells. Toxicol Appl Pharmacol. 338:124-133.
Wei H et. al. (January 2017). Role of oxidative stress and DNA hydroxymethylation in the neurotoxicity of fine particulate matter.Toxicology.
Kowluru RA et. al. (July 2016). Dynamic DNA methylation of matrix metalloproteinase-9 in the development of diabetic retinopathy.Lab Invest.
Celarain N et. al. (April 2016). TREM2 upregulation correlates with 5-hydroxymethycytosine enrichment in Alzheimer's disease hippocampus. Clin Epigenetics. 8:37.
Xu X et. al. (March 2015). Epigenetic balance of aberrant Rasal1 promoter methylation and hydroxymethylation regulates cardiac fibrosis. Cardiovasc Res. 105(3):279-91.
Tampe B et. al. (December 2014). Induction of Tet3-dependent Epigenetic Remodeling by Low-dose Hy. EBioMedicine. 1(1)
Bellizzi D et. al. (December 2013). The control region of mitochondrial DNA shows an unusual CpG and non-CpG methylation pattern. DNA Res. 20(6):537-47.
Hatem K. Abbas et. al. Detection of siRNA-Directed DNA Methylation and Gene Silencing in Response to Salinity Stress Iraqi Academic Scientific Journals. 16(2):89-110.
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 |
| DNA羟甲基化免疫共沉淀试剂盒 |
24 次 |
英文PDF--中文PDF |
A-P-1038-24 |
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| DNA羟甲基化免疫共沉淀试剂盒 |
48 次 |
英文PDF--中文PDF |
A-P-1038-48 |
|
| DNA羟甲基化免疫共沉淀试剂盒 |
96 次 |
英文PDF--中文PDF |
A-P-1038-96 |
|
表观遗传学产品全面解决方案列下：
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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。 |
