胸腺嘧啶DNA糖基化酶(TDG)活性/抑制分析试剂盒(比色法)                                                                  货号：A-P-3094

Epigenase^TM Thymine DNA Glycosylase (TDG) Activity/Inhibition Assay Kit (Colorimetric) 

背景资料：DNA demethylation has been demonstrated to occur through at least two different pathways: (1) active deamination of 5-mC to T by an AID/APOBEC enzyme, giving a G/T mismatch that is converted to G/C by thymine DNA glycosylase (TDG) and then subsequently base excision repair (BER); and (2) iterative oxidation of 5-mC by TET enzyme to 5-hmC and further to 5-formycytosine (5-fC) and 5-carboxylcytosine (5-caC). Both 5-fC and 5-caC can be rapidly excised by TDG to allow subsequent BER processing which results in 5-fC and 5-caC converting back to unmodified cytosine.

TDG belongs to the TDG/mug DNA glycosylase family. Besides playing a critical role in active DNA methylation, TDG also removes thymine moieties from G/T mismatches by hydrolyzing the carbon-nitrogen bond between the sugar-phosphate backbone of DNA and the mismatched thymine. It has been shown that TDG participates in the regulation of embryonic and germ cell development, mediates cellular defense against genetic and epigenetic mutations, and mediates DNA-directed cytotoxicity. In addition, TDG is a proposed tumor suppressor candidate. Thus, the measurement of TDG activity would be important as TDG may be involved in preventing tumorigenesis and determining cellular response to chemotherapy through DNA demethylation and BER, thereby benefiting cancer diagnostics and new target-based cancer therapeutics. 

The currently used methods for TDG activity detection involve DNA cleavage followed by electrophoresis, which can be time consuming and inconvenient, with low throughput and high cost. To address this issue, Epigentek developed and offers the Epigenase™ Thymine DNA Glycosylase (TDG) Activity/Inhibition Assay Kit (Colorimetric).

产品描述：In this assay, a 5-fC DNA substrate is stably coated onto microplate wells. Active TDG binds to the substrate and excises 5-fC from the substrate by a nicking reaction. The remaining un-excised 5-fC in the substrate will be recognized with a high affinity 5-fC DNA antibody. The ratio or amount of 5-fC in the substrate, which is inversely proportional to enzyme activity, can then be colorimetrically measured by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The activity of the TDG enzyme is in turn inversely proportional to the optical density intensity measured.The Epigenase™ Thymine DNA Glycosylase (TDG) Activity/Inhibition Assay Kit (Colorimetric) is a complete set of optimized buffers and reagents for measuring the activity/inhibition of total TDG enzyme in nuclear extracts or purified TDG enzymes from a broad range of species such as mammals, plants, fungi, and bacteria, in a variety of forms including but not limited to cultured cells and fresh and frozen tissues.

产品特点：

1·Colorimetric assay with easy-to-follow steps for convenience and speed. The entire procedure can be finished within 5 hours.  比色法检测流程简单易用，方便，快速，完成时间需要5小时 

2·Directly measures TDG activity without the need for DNA cleavage, electrophoresis, or chromatography.方便的直接测定TDG活性，无需DNA裂解，电泳或色谱分析；

3·Both cell/tissue extracts and purified TDG proteins can be used, which allows for detection of inhibitory effects of TDG inhibitors in vivo and in vitro.可以使用细胞/组织核提取物和纯化的TDG蛋白，容许在活的有机体内或体外检测TDG的抑制活性；

4·Novel assay principle allows high sensitivity to be achieved. The activity can be detected from as low as 20 ng of purified TDG protein.高灵敏度，其活性检测可低至20ng纯化的TDG蛋白；

5·TDG assay standard is conveniently included, which allows for specific quantification of TDG activity.本试剂盒包含的标准品，能特异的定量TDG的活性；

6·Strip microplate format makes the assay flexible: manual or high throughput analysis (96 assays).微孔板形式即适用于手工操作又适用于高通量分析（96次实验）。

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
胸腺嘧啶DNA糖基化酶(TDG)活性/抑制分析试剂盒(比色法)	48 次	PDF	A-P-3094-48
胸腺嘧啶DNA糖基化酶(TDG)活性/抑制分析试剂盒(比色法)	96 次	PDF	A-P-3094-96

表观遗传学产品全面解决方案列下：

产品名称	规格	操作手册	货号	询价
DNA羟甲基化定量试剂盒(荧光法)	48 次	PDF	A-P-1037-48
DNA羟甲基化定量试剂盒(荧光法)	96 次	PDF	A-P-1037-96
DNA羟甲基化定量试剂盒(比色法)	48 次	PDF	A-P-1036-48
DNA羟甲基化定量试剂盒(比色法)	96 次	PDF	A-P-1036-96
DNA甲基化定量试剂盒(荧光法)	48 次	PDF	A-P-1035-48
DNA甲基化定量试剂盒(荧光法)	96 次	PDF	A-P-1035-96
DNA甲基化定量试剂盒(比色法)	48 次	PDF	A-P-1034-48
DNA甲基化定量试剂盒(比色法)	96 次	PDF	A-P-1034-96
通用DNA甲基化定量试剂盒	48 次	PDF	A-P-1011-48
通用DNA甲基化定量试剂盒	96 次	PDF	A-P-1011-96
超敏DNA甲基化定量试剂盒	48 次	PDF	A-P-1021-48
超敏DNA甲基化定量试剂盒	96 次	PDF	A-P-1021-96
总DNA甲基化定量试剂盒	48 次	PDF	A-P-1014-48
总DNA甲基化定量试剂盒	96 次	PDF	A-P-1014-96

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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