Jones I, Madej A
Department of Clinical Chemistry, College of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
J Immunoassay. 1988;9(3-4):349-65. doi: 10.1080/01971528808053221.
Microtitre plates were coated with antiserum against oestradiol-17 beta-6-(O-carboxymethyl)-oxime bovine serum albumin raised in sheep. The plasma samples (0.2-1.0 ml) were extracted with peroxide-free diethyl ether prepared daily by treatment with Al2O3. The enzyme conjugate was prepared by coupling oestradiol-17 beta-6-(O-carboxymethyl)-oxime to horse-radish peroxidase. The conjugate was chromatographed on a Sephadex G-25 column. The standard curve ranged from 0.37 to 18.40 fmol/well of oestradiol-17 beta. The amount of oestradiol-17 beta causing a 50% reduction of maximum binding was 4.4 fmol/well. Standards and samples were incubated overnight at 4 degrees C. The conjugate solution was added followed by further incubation for 2 h at 4 degrees C. Tetramethylbenzidine was used as a chromogen, and the optical density was measured at 450 nm. The patterns of oestradiol-17 beta during a normal oestrus cycle in the cow and mare are presented.
微量滴定板用羊抗17β-雌二醇-6-(O-羧甲基)-肟牛血清白蛋白抗血清包被。血浆样本（0.2 - 1.0毫升）用每天用氧化铝处理制备的无过氧化物二乙醚萃取。酶结合物通过将17β-雌二醇-6-(O-羧甲基)-肟与辣根过氧化物酶偶联制备。结合物在葡聚糖凝胶G - 25柱上进行色谱分析。标准曲线范围为每孔0.37至18.40飞摩尔的17β-雌二醇。导致最大结合减少50%的17β-雌二醇量为每孔4.4飞摩尔。标准品和样本在4℃下孵育过夜。加入结合物溶液，然后在4℃下进一步孵育2小时。使用四甲基联苯胺作为显色剂，并在450纳米处测量光密度。给出了母牛和母马正常发情周期中17β-雌二醇的模式。
