大鼠血清中褪黑素的放射免疫测定

Lee J. Grota , Victor Snieckus , S.Osmond De Silva , Hing W. Tsui , William R. Holloway , Alfred J. Lewy , Gregory M. Brown
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引用次数: 50

摘要

1.1. 褪黑素通过na -对羧基苯基、na -丙酸、亚甲基或重氮化对氨基苯甲酸桥接与白蛋白偶联,注射到兔或羊体内。所有偶联过程均能刺激与褪黑素结合最密切的抗血清;n -乙酰5 -羟色胺和6-羟褪黑素在3-5%时与抗血清发生交叉反应。其他天然存在的吲哚烷基胺衍生物表现出较少的交叉反应性。通过在吲哚N上或附近的桥接将褪黑素半抗原与蛋白质偶联可以刺激对褪黑素具有相对特异性的抗血清,这表明使用这些偶联过程可以产生与任何其他单个吲哚烷基胺衍生物特异性结合的抗血清。利用亚甲基桥接抗血清,开发并验证了大鼠血清中褪黑素的放射免疫测定。除了可靠性和平行性测试外,血清中褪黑素的放射免疫测定与气相色谱-质谱测定的相关性为0.98。血清褪黑素水平有一个典型的24小时节律,在黑暗期后期达到峰值(L:D 12:12)。注射50微克褪黑素后,血清浓度达到生理水平的5-20倍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Radioimmunoassay of melatonin in rat serum

  • 1.

    1. Melatonin was coupled to albumin by means of a Na-p-carboxybenzyl, a Na-propionic acid, methylene, or a diazotized p-aminobenzoic acid bridge and injected into rabbits or sheep.

  • 2.

    2. All coupling procedures stimulated antisera which bound melatonin with the greatest affinity; N-acetylserotonin and 6-hydroxymelatonin cross-reacted with the antisera at 3–5%. Other naturally occurring indolealkylamine derivatives showed less cross-reactivity.

  • 3.

    3. Coupling the hapten melatonin to protein through a bridge at or near the indole N can stimulate antisera which are relatively specific to melatonin and suggest that antisera which bind specifically to any other individual indolealkylamine derivative could be produced using these coupling procedures.

  • 4.

    4. Using the methylene bridged antisera, a radioimmunoassay for melatonin was developed and validated for rat serum. In addition to tests for reliability and parallelism, serum estimates of melatonin by radioimmunoassay correlated .98 with gas chromatography-mass spectrometry.

  • 5.

    5. Serum melatonin levels have a characteristic 24-hour rhythm with a crest late in the dark period (L:D 12:12). Following the injection of 50 ug of melatonin, serum levels reach 5–20 fold higher concentrations than physiological levels.

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