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使用 N 端甲状旁腺激素抗血清的血浆甲状旁腺激素(PTH)放射免疫测定法(作者译)

[A radioimmunoassay for plasma parathyroid hormone (PTH) using N-terminal PTH antiserum (author's transl)].

作者信息

Kayamori R, Yamada Y, Ito S, Iwasaki Y, Hayashi M, Momotu T, Takai K, Miyashita M, Kaneko K, Shibata A, Nara Y, Suzuki M, Hirasawa Y

出版信息

Nihon Naibunpi Gakkai Zasshi. 1979 Nov 20;55(11):1372-83. doi: 10.1507/endocrine1927.55.11_1372.

Abstract

In order to investigate plasma bioactive PTH, we tried to assay the N-terminal portion of PTH by RIA. The antiserum to PTH was prepared by immunizing rabbits with a bovine 1-34 PTH conjugate BSA. A preparation of labeled PTH was radioiodinated by the chloramine-T or lactoperoxidase method. Labeled PTH was purified by means of adsorption by Quso G-32 powder or a sephadex G-50. The separation of the free and bound labeled hormone was performed by the dextran-coated charcoal method. The assay was carried out as follows: 0.2 ml diluted buffer (0.05 M, pH 8.6, veronal buffer), 0.1 ml standard PTH or sample to be tested, and 0.1 ml anti-PTH serum were mixed. After the first incubation at 4 degrees C for 4 days, 0.1 ml labeled PTH were added. After a second incubation at 4 degrees C for 12 hours, the assay tubes were centrifuged at 2,000 rpm for 30 min and the precipitates were counted. Various hypothalamic, pituitary and thyroid hormones did not interfere with the RIA for PTH. A dose response curve was obtained in a range from 100 pg to 5,000 pg per ml of standard PTH in this assay system. The serum immunoreactive PTH in healthy subjects values less than 290 pg per ml.

摘要

为了研究血浆生物活性甲状旁腺激素(PTH),我们尝试用放射免疫分析法(RIA)检测PTH的N端部分。用牛1 - 34 PTH与牛血清白蛋白(BSA)的偶联物免疫兔子制备PTH抗血清。通过氯胺 - T法或乳过氧化物酶法对PTH制剂进行放射性碘化标记。标记的PTH通过Quso G - 32粉末或葡聚糖凝胶G - 50吸附进行纯化。采用葡聚糖包被活性炭法分离游离和结合的标记激素。检测方法如下:将0.2 ml稀释缓冲液(0.05 M,pH 8.6,巴比妥缓冲液)、0.1 ml标准PTH或待测样品以及0.1 ml抗PTH血清混合。在4℃首次孵育4天后,加入0.1 ml标记的PTH。在4℃再次孵育12小时后,将检测管以2000转/分钟的速度离心30分钟并对沉淀物进行计数。各种下丘脑、垂体和甲状腺激素不干扰PTH的放射免疫分析。在该检测系统中,标准PTH浓度在每毫升100 pg至5000 pg范围内获得了剂量反应曲线。健康受试者血清免疫反应性PTH值低于每毫升290 pg。

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