Aranda Iriarte A M
Reproduccion. 1980 Apr-Jun;4(2):109-33.
A rat LH radioimmunoassay (RIA) using the immunoreactants provided by Dr Parlow on behalf of the Rat Pituitary Hormone Program of the NIAMDD of the NIH is described. The instructions suggested by the NIH have been modified in order to reach a higher sensitivity. It is possible to use prepipetted frozen standards and the same labelled antigen over a period of 1 month, or longer, after labelling, provided the preparation is purified immediately prior to its use. These procedures, as well as the addition of antibody and buffer simultaneously, shorten considerably the time spent on a given RIA and decrease the inter- and intra-assay variability. The specificity of the method is also described. There is good parallelism between the standard curve and serial dilutions of plasmas and hypophyses of rats. The RIA was validated physiologically by LH determination on plasmas obtained from animals submitted to different experimental situations in which a decrease (hypophysectomy) or increase (ovariectomy, LH-RH treatment, etc.) of this hormone is expected.
本文描述了一种使用帕洛博士代表美国国立卫生研究院(NIH)国家糖尿病、消化和肾脏疾病研究所(NIAMDD)大鼠垂体激素项目提供的免疫反应物进行大鼠促黄体生成素(LH)放射免疫分析(RIA)的方法。对NIH建议的操作说明进行了修改,以提高灵敏度。可以使用预先分装的冷冻标准品,并且标记后的同一标记抗原在标记后1个月或更长时间内均可使用,前提是在使用前立即对制剂进行纯化。这些操作程序,以及同时加入抗体和缓冲液,大大缩短了给定RIA所需的时间,并降低了批间和批内变异。还描述了该方法的特异性。标准曲线与大鼠血浆和垂体的系列稀释液之间具有良好的平行性。通过对处于不同实验情况下动物的血浆进行LH测定,从生理学角度验证了该RIA,在这些实验情况下,预期该激素会减少(垂体切除)或增加(卵巢切除、LH-RH治疗等)。