| Quantification of Uric Acid or Xanthine in Plant Samples
|
|
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）
|
|
|
| Extraction of Total Proteins from Rice Plant
|
|
Author:
Date:
2014-11-05
[Abstract] This protocol provides an efficient method for preparation of high-quality proteins from rice leaves and grains. The method involves phenol extraction to separate proteins from the non-protein components such as polysaccharides, lipids and phenolic compounds that are commonly enriched in plant tissues. Following isolation, proteins are precipitated with ammonium acetate/methanol and then solubilized for proteomic analysis. As the protocol is simple, universal, and most importantly compatible with silver staining, it has been applied to our routine protein extraction from rice and many other plant tissues and it even works fine in animal tissues for the requirement of electrophoretic separation.
[摘要] 这个协议提供了一种从稻叶和谷物制备高质量蛋白质的有效方法。 该方法包括酚提取以从通常富含植物组织的非蛋白质组分如多糖，脂质和酚类化合物中分离蛋白质。 分离后，用乙酸铵/甲醇沉淀蛋白质，然后溶解用于蛋白质组分析。 由于协议简单，通用，最重要的是与银染色兼容，它已经应用于我们从水稻和许多其他植物组织的常规蛋白质提取，甚至在动物组织中对于电泳分离的要求很好。
|
|
|