Furlow J D, Ahrens H, Mueller G C, Gorski J
Department of Biochemistry, University of Wisconsin, Madison 53706.
Endocrinology. 1990 Sep;127(3):1028-32. doi: 10.1210/endo-127-3-1028.
Specific polyclonal antisera to the rat estrogen receptor (rER) were developed using a synthetic peptide as the antigen. The peptide corresponds to amino acids 270-284 deduced from the cloned rER cDNA and has no homology to other steroid hormone receptors. Antipeptide antisera raised in rabbits specifically recognize a 67,000 mol wt protein, shown to be the rER, in Western blot experiments. In immunoprecipitation experiments, one tested antiserum bound unoccupied as well as 17 beta-estradiol-occupied rERs, indicating that this region is exposed in both receptor forms. This antiserum shows no cross-reactivity with rat progesterone or glucocorticoid receptors. Cross-reactivity with rabbit, human, and, to a lesser extent, bovine ERs was observed.
使用合成肽作为抗原制备了针对大鼠雌激素受体(rER)的特异性多克隆抗血清。该肽对应于从克隆的rER cDNA推导的氨基酸270 - 284,与其他类固醇激素受体无同源性。在兔中产生的抗肽抗血清在蛋白质印迹实验中特异性识别一种67,000分子量的蛋白质,证明其为rER。在免疫沉淀实验中,一种测试抗血清能结合未被占据以及被17β-雌二醇占据的rER,表明该区域在两种受体形式中均暴露。这种抗血清与大鼠孕酮或糖皮质激素受体无交叉反应。观察到与兔、人以及程度较轻的牛ER有交叉反应。