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-2 (4-carboxy phenyl)-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide


Company: Sigma-Aldrich
Catalog#: C221
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Localisation and Quantification of Reactive Oxygen Species and Nitric Oxide in Arabidopsis Roots in Response to Fungal Infection
[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。