| Simple Spectroscopic Determination of Nitrate, Nitrite, and Ammonium in Arabidopsis thaliana
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Author:
Date:
2017-05-20
[Abstract] Plants use nitrate, nitrite, and ammonium as inorganic nitrogen (N) sources. These N compounds are included in plant tissues at various concentrations depending on the balance between their uptake and assimilation. Thus, the contents of nitrate, nitrite, and ammonium are physiological indicators of plant N economy. Here, we describe a protocol for measurement of these inorganic N species in A. thaliana shoots or roots.
[摘要] 植物使用硝酸盐，亚硝酸盐和铵作为无机氮（N）源。这些N化合物根据其吸收和同化之间的平衡以各种浓度包含在植物组织中。因此，硝酸盐，亚硝酸盐和铵的含量是植物N经济的生理指标。在这里，我们描述了在A中测量这些无机N物质的方案。 thaliana 芽或根。
背景 无机氮含量的测定对于预测植物吸收和吸收氮的能力很重要。研究人员经常使用需要昂贵设备的技术，如高效液相色谱（HPLC），用于这些测量。本协议基于通用光谱法与廉价的试剂，使其适用于许多研究人员。在酸性条件下，硝酸盐引起水杨酸的硝化，随后加入碱性溶液会导致黄色复合物。在酸性条件下，亚硝酸盐与磺胺类反应产生重氮化合物，其与N - （1-萘基）乙二胺进行重氮偶联，形成粉红色偶氮化合物。在硝普钠盐的催化作用下，铵可以测定为蓝色靛酚衍生物。每个实验程序容易，快速，简单。
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| Signaling Assays for Detection of Human G-protein-coupled Receptors in Yeast
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Author:
Date:
2014-08-20
[Abstract] G-protein-coupled receptors (GPCRs) are the largest group of cell-surface proteins and are major molecular targets for drug development. The protocol described herein is for the detection of human GPCR signaling in the yeast Saccharomyces cerevisiae. Using Zoanthus sp. green fluorescent protein (ZsGreen) as the reporter, engineered yeast cells expressing human GPCRs emit strong fluorescence in response to stimuli leading to receptor signal activation. This assay method would allow screening for agonistic ligands and critical mutations required for human GPCR signaling.
[摘要] G蛋白偶联受体（GPCR）是最大的细胞表面蛋白质组，是药物开发的主要分子靶标。 本文所述的方案用于在酵母酿酒酵母中检测人GPCR信号传导。 使用 Zoanthus 绿色荧光蛋白（ZsGreen ）作为报告物，表达人GPCR的工程化酵母细胞响应于导致受体信号活化的刺激而发射强荧光。 该测定方法将允许筛选激动剂配体和人GPCR信号传导所需的关键突变。
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