Radioimmunoassay for Rat Prolactin∗
Radioimmunoassay for Rat Prolactin∗
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大鼠催乳素放射免疫测定＊
DOI：
10.3181/00379727-130-33657
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发表时间：
1969
影响因子：
--
通讯作者：
S. Ellis
中科院分区：
文献类型：
--
作者：
G. Niswender;C. L. Chen;A. Midgley;J. Meites;S. Ellis
测定生物样品中催乳素最常用的方法是里昂鸽子作物生物测定法(1)。用这种方法可以估计垂体制剂和含有这种激素相对高浓度的其他组织的催乳素含量（至少约0.03 IU或2 μg的NIH-P-B1, 13 IU/mg）；然而，不可能测定正常动物血清中的催乳素。最近，针对绵羊(2)和牛催乳素(3)的双抗体放射免疫测定技术已经被开发出来，而针对大鼠催乳素(4)的放射免疫测定技术也有报道，该技术使用色谱电泳步骤从抗体结合的游离催乳素-125 I中分离出来。这些技术对定量测定血清催乳素水平有足够的敏感性。本通信描述了一种大鼠催乳素双抗体放射免疫测定法的发展，该方法能够测量100 μl未成熟雌性大鼠血清或25 μl正常雌性大鼠发情期间血清中的催乳素含量。方法与材料。纯化的大鼠催乳素（HIV-8-C）用于放射碘和免疫，由Ellis等人从整个冷冻大鼠脑垂体中制备(5)。在本研究的其余部分中，该制剂被命名为B329。通过生理操作三组成熟雌性Sprague-Dawley大鼠获得其他具有不同浓度大鼠催乳素的制剂。第一组大鼠（B350）切除卵巢，每日皮下注射含苯甲酸雌二醇10 μg的玉米油，连续10 d。第二组（B349）切除卵巢后注射玉米油10 d，第三组（B351）为未处理的正常雌性大鼠。2周后经腹腔静脉终末放血，将各组血汇集，离心取血清。
The most commonly used method for measuring prolactin in biological samples has been the pigeon crop bioassay of Lyons (1). It is possible with this method to estimate the prolactin content of pituitary preparations and of other tissues containing relatively high concentrations of this hormone (a minimum of about 0.03 IU or 2 μg of NIH-P-B1, 13 IU/mg); however, it was not possible to assay the prolactin present in serum from normal animals. Recently, double antibody radioimmunoassay techniques have been developed for ovine (2) and bovine prolactin (3) and a radioimmunoassay for rat prolactin (4) which uses a chromatoelectrophoresis step to separate free prolactin-125 I from that bound to the antibody has also been reported. These techniques are sufficiently sensitive to quantitate serum levels of prolactin. This communication describes the development of a double antibody radioimmunoassay for rat prolactin capable of measuring the amount of prolactin in 100 μl of serum from immature female rats or in 25 μl of serum from normal females during diestrus. Methods and Materials. Purified rat prolactin (HIV-8-C) for radioiodination and for immunization was prepared from whole, frozen rat pituitary glands by Ellis et al. (5). This preparation is designated B329 in the remainder of the present study. Other preparations with differing concentrations of rat prolactin were obtained by physiologic manipulation of three groups of mature, female Sprague-Dawley rats. Rats in the first group (B350) were ovariectomized and injected subcutaneously with 10 μg of estradiol benzoate in corn oil daily for 10 days. Rats in the second group (B349) were ovariectomized and injected with corn oil for 10 days while the third group was composed of untreated intact females during diestrus (B351). At the end of 2 weeks the rats were bled terminally via the abdominal vena cava, and the blood from each group was pooled and serum was obtained by centrifugation.