A Homologous Radioimmunoassay for Bullfrog Basic Gonadotropin
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概要
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A double antibody radioimmunoassay (RIA) for the measurement of bullfrog basic gonadotropin (GTH) is described. The antiserum was raised against a highly purified bullfrog basic GTH. Bullfrog basic GTH-IV (pI=9.33) was used for radioiodination and as a reference standard. The dissociation constant of the binding of the antibody to bullfrog basic GTH-IV was 8.28×10<SUP>-12</SUP>M. The standard curve was obtained with 0.1-25.6ng/ml of bullfrog basic GTH-IV. When bullfrog sera were assayed by this RIA system, the intra-assay coefficient of variation and inter-assay coefficient of variation were 4.45% and 5.6%, respectively. The accuracy of the assay was shown as the recovery of bullfrog basic GTH-IV added to the pooled bullfrog sera. With a range of 0.1-12.8ng/ml, the recovery was 104.78±11.91%(Mean±S. D.). When bullfrog pituitary extract was fractionated by isoelectric focusing (IEF) and assayed for gonadotropin by this RIA system and also by radioreceptor assay systems employing <I>Xenopus</I> or Anolis testicular homogenates, the RIA positive peaks agreed well with those obtained with <I>Xenopus</I> RRA, indicating that the present RIA system measures LH-like gonadotropin. The inhibition curves of pituitary preparations from bullfrog, brown frogs and dark-colored frog were almost parallel to the standard curve, while those of other anuran and urodele species were not. This RIA system was highly sensitive, but allowed only a narrow application, because of species-specificity.
- 社団法人 日本内分泌学会の論文
著者
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TANAKA SHIGEYASU
Institute of Endocrinology, Gunma University
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Wakabayashi Katsumi
Institute For Molecular And Cellular Regulation Gunma University
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Tanaka Shigeyasu
Institute Of Endocrinology Gunma University
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Hanaoka Yoichi
Institute Of Endocrinology Gunma University
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HANAOKA YOUICHI
Institute of Endocrinology, Gunma University
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