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首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >A nonradioactive iodide uptake assay for sodium iodide symporter function
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A nonradioactive iodide uptake assay for sodium iodide symporter function

机译:碘化钠共转运蛋白功能的非放射性碘吸收测定

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摘要

The standard assay for sodium iodide symporter (NIS) function is based on the measurement of radioiodide uptake (~(125)I) in NIS-expressing cells. However, cost and safety issues have limited the method from being used widely. Here we describe a simple spectrophotometric assay for the determination of iodide accumulation in rat thyroid-derived cells (FRTL5) based on the catalytic effect of iodide on the reduction of yellow cerium(IV) to colorless cerium(III) in the presence of arsenious acid (Sandell-Kolthoff reaction). The assay is fast and highly reproducible with a Z′ factor of 0.70. This procedure allows the screening of more than 800 samples per day and can easily be adapted to robotic systems for high-throughput screening of NIS function modulators. Using this method, the potency of several known inhibitors of NIS function was evaluated in a single day with high accuracy and reliability. Measured IC_(50) values were essentially identical to those determined using Na~(125)I.
机译:碘化钠共转运蛋白(NIS)功能的标准检测方法是基于表达NIS的细胞中放射性碘吸收(〜(125)I)的测量。但是,成本和安全问题限制了该方法的广泛使用。在此,我们描述了一种简单的分光光度法,该方法用于根据砷在砷酸存在下将黄色铈(IV)还原为无色铈(III)的催化作用,确定大鼠甲状腺衍生细胞(FRTL5)中碘的积累。 (Sandell-Kolthoff反应)。该测定快速且高度可重复,Z'因子为0.70。该程序每天可以筛选800多个样品,并且可以轻松地应用于机器人系统,以对NIS功能调节剂进行高通量筛选。使用这种方法,可以在一天之内以高精度和可靠性评估几种已知的NIS功能抑制剂的效力。测得的IC_(50)值与使用Na〜(125)I测得的值基本相同。

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