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H2DCFDA (H2-DCF, DCF)

H2DCFDA(H2-DCF,DCF)

Company: Thermo Fisher Scientific
Catalog#: D399
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Detection of Reactive Oxygen Species in Oryza sativa L. (Rice)
Author:
Date:
2016-12-20
[Abstract]  Superoxide ions (O2-) and hydrogen peroxide (H2O2) are the reactive oxygen species (ROS) that play a significant role in regulation of many plant processes. The level of O2- ions is determined qualitatively using nitrobluetetrazolium (NBT) assay while the H2O2 is qualitatively estimated using 3,3-diaminobenzidine (DAB) and 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFDA) assay. Further the aqueous content of H2O2 is estimated quantitatively using ferrous oxidation-xylenol orange (FOX) assay. [摘要]  超氧化物离子(O 2 - )和过氧化氢(H 2 O 2 O 2)是起重要作用的活性氧(ROS)在许多植物过程的调节。使用硝基四氮唑(NBT)测定定性地测定O 2 - 离子的水平,同时使用3,3 - 二氨基联苯胺(DAB)和2',7'-二氯二氢荧光素二乙酸酯(H 2 N 2 DCFDA)测定。此外,使用亚铁氧化 - 二甲苯酚橙(FOX)测定定量地估计H 2 O 2 O 2的含水量。

背景 超氧化物离子和过氧化氢是在许多情况下起主要作用的活性氧分子涉及植物生长发育的过程包括非生物胁迫耐受性。为了更好地了解ROS对这些过程的调节,对不同类型ROS的定性和定量估计具有重要意义。通过由NADPH氧化酶系统介导的电子从NADPH转移到氧(O 2),产生O 2。使用NBT测定法在水稻幼苗中估计这些离子,其基于通过O将黄色NBT还原成深蓝色不溶性甲the的原理(Kaur等人,2016)。
  H 2是另一种作为调节不同植物过程的重要信号分子的活性氧分子。在使用DAB和H 2 DCFDA测定的水稻幼苗中定性地估计H 2 O 2 O 2的含量(Kaur等人,2016)。 DAB测定是基于DAB与H 2 O 2 O 2的反应形成深棕色聚合产物的原理,而H 2 N 2 ...

Localisation and Quantification of Reactive Oxygen Species and Nitric Oxide in Arabidopsis Roots in Response to Fungal Infection
Author:
Date:
2014-10-05
[Abstract]  Nitric oxide and reactive oxygen species have emerged as important signalling molecules in plants. The half-lives of NO and ROS are very short therefore rapid and precise measurements are required for the understanding biological roles of these redox active species. Various organelles and compartments generate NO and ROS thus it is important to determine precise location of these free radicals in order to understand their signalling roles. Diaminofluorescen (DAF) and fluorescent 2', 7'-dichlorofluorescein (DCF) dyes are employed to determine NO and ROS localisation. The advantage of this approach is that the dyes diffuse precisely to NO and ROS producing sites and generate fluorescence which can be detected by fluorescence- or confocal laser scanning microscopes. However, this technique ... [摘要]  一氧化氮和活性氧在植物中作为重要的信号分子出现。 NO和ROS的半衰期非常短,因此需要快速和精确的测量来理解这些氧化还原活性物质的生物学作用。各种细胞器和隔室产生NO和ROS,因此重要的是确定这些自由基的精确位置,以了解他们的信号传导作用。使用二氨基荧光素(DAF)和荧光2',7'-二氯荧光素(DCF)染料来确定NO和ROS定位。这种方法的优点是染料精确扩散到NO和ROS产生位点,并产生可以通过荧光或共聚焦激光扫描显微镜检测的荧光。然而,这种技术有其缺点;特别是需要建立荧光信号的特异性。因此,需要诸如cPTIO和抗坏血酸的ROS的使用清除剂来确认荧光信号的特异性,并且理想地,由于与每种方法相关的优点和缺点,理想地确认使用其它方法获得的数据(Gupta和Igamberdiev,2013 )。在这里我们描述了一种方法来检测响应于木霉属,镰刀菌属使用DAF,气相Griess试剂测定和DCF荧光方法的感染的拟南芥根的NO和ROS产生。

Flow Cytometric Detection of Reactive Oxygen Species
Author:
Date:
2013-04-20
[Abstract]  Reactive oxygen species (ROS) are molecules containing hydroxyl radicals or peroxides with unpaired electrons. In healthy aerobic cells, ROS are produced naturally as a byproduct of oxidative phosphorylation, oxidoreductase enzymes, or metal catalyzed oxidation at a controlled rate. However, ROS can be induced under some stress conditions especially exposure to environmental oxidants and certain drugs that leads to oxidative stress. Exceed ROS can cause damages in the building blocks of cells including DNA, proteins, and lipids, and eventually results in cell death. Cell-permeant 2', 7'-dichlorodihydrofluorescein diacetate (H2DCFDA) is a widely used ROS indicator. The reduced non-fluorescent fluorescein H2DCFDA can be oxidized and converted into fluorescent 2’, ... [摘要]  活性氧(ROS)是含有羟基自由基的分子或具有不成对电子的过氧化物。 在健康好氧细胞中,ROS作为氧化磷酸化,氧化还原酶或金属催化氧化的副产物以受控速率天然产生。 然而,ROS可以在一些应激条件下诱导,特别是暴露于环境氧化剂和导致氧化应激的某些药物。 Exceed ROS可导致细胞构建块(包括DNA,蛋白质和脂质)的损伤,并最终导致细胞死亡。 细胞渗透剂2',7'-二氯二氢荧光素二乙酸酯(H 2 DCFDA)是广泛使用的ROS指示剂。 还原的非荧光荧光素H 2 DCFDA可以被氧化并通过细胞内ROS转化为荧光2',7'-二氯荧光素(DCF)。 在该协议中,我们应用H 2 DCFDA标记细胞内ROS并通过流式细胞术检测DCF强度。

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