| Quantification of Uric Acid or Xanthine in Plant Samples
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Author:
Date:
2015-07-05
[Abstract] We developed this protocol to assay and quantify the content of uric acid or xanthine in various tissues of Arabidopsis thaliana mutant lines with defective urate oxidase or xanthine dehydrogenase1 and in their complementation and suppressor lines (Hauck et al., 2014).
The protocol is based on a method developed by Invitrogen Life Technologies for measuring uric acid or xanthine in human serum (see References 2 and 3). That protocol though required two adaptions for its use in plant science. Firstly by heating the plant samples, the activity of urate oxidase and xanthine dehydrogenase in the wild type samples is eliminated. Wild type extracts always serve as the proper pigmentation background when calculating the standard curves of uric acid and xanthine. ...
[摘要] 我们开发了该方案以测定和定量具有缺陷的尿酸氧化酶或黄嘌呤脱氢酶1的拟南芥突变体系以及它们的互补和抑制系的各种组织中的尿酸或黄嘌呤的含量（Hauck等</em>，2014）。
该方案基于由Invitrogen Life Technologies开发的用于测量人血清中的尿酸或黄嘌呤的方法（参见参考文献2和3）。该协议虽然需要两个适应它在植物科学中的使用。首先通过加热植物样品，消除野生型样品中的尿酸氧化酶和黄嘌呤脱氢酶的活性。当计算尿酸和黄嘌呤的标准曲线时，野生型提取物总是作为适当的色素沉着背景。其次，在添加和不添加尿酸氧化酶或黄嘌呤脱氢酶的情况下测量所有样品，以校正由先前应激诱导的样品中的任何H 2 O 2 O 2。 >该测定基于以下一对偶联反应：</1> 1）尿酸+ O 2→羟基香草酸+ H 2 O 2 - （辣根过氧化物酶反应）的反应物（尿酸氧化酶反应）的反应。 ）
因此对于黄嘌呤：
1）黄嘌呤+ H 2 O + O 2→尿酸+ H 2 - O 2（黄嘌呤氧化酶反应）2→AR + H 2→O→2→Resorufin + O 2→ （辣根过氧化物酶反应）
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| Glycolate Oxidase Activity Assay in Plants
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Author:
Date:
2012-10-20
[Abstract] Glycolate oxidase is located in the peroxisome and is involved in the photorespiratory cycle which recovers some of the carbon loss during photosynthesis. Glycolate oxidase converts glycolate to glyxoylate with the concomitant production of H2O2.In this assay, the H2O2 generated, in the presence of HRP, oxidizes O-dianisidine into a colored O-dianisidine radical cation that can be quantified spectrophotometrically. The amount of color produces is directly proportional to the glycolate oxidase activity.
[摘要] 乙醇酸氧化酶位于过氧化物酶体中并且参与光呼吸循环，其恢复光合作用期间的一些碳损失。 乙醇酸氧化酶将乙醇酸转化为乙醛酸，同时产生H 2 O 2 Sub。在该测定中，H 2 O 2 O 2 - 在HRP的存在下，将O-联茴香胺氧化成可以用分光光度法定量的有色的O-联苯胺自由基阳离子。 产生的颜色量与乙醇酸氧化酶活性成正比。
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