Dicker P D, Tsai S Y, Weigel N L, Tsai M J, Schrader W T, O'Malley B W
J Steroid Biochem. 1984 Jan;20(1):43-50. doi: 10.1016/0022-4731(84)90187-0.
The progesterone receptor of the hen oviduct is composed of two non-identical hormone-binding polypeptide subunits, A (Mr = 79,000) and B (Mr = 108,000). We used a highly purified preparation of B to immunize mouse spleen cells in vitro. After 5 days in culture, the cells were fused with SP2/0-Ag 14 myeloma cells. The resultant hybridomas were screened using an enzyme linked immunosorbent solid phase assay, and those hybridomas producing antibodies binding to the immunogen were cloned by limiting dilution. One such clone, 9B3-12, secreted an antibody of immunoglobulin class IgM, which binds to B. This was indicated by the ability of the antibody to increase the rate of sedimentation coefficient of the B subunit. Further, when the proteins in the B preparation were separated by electrophoresis and blotted onto nitrocellulose filters the antibody bound to a protein of 108,000 daltons. The antibody produced by 9B3-12 also reacted with subunit A and with the human progesterone receptor but failed to bind to the chick liver glucocorticoid receptor or to progesterone in the absence of its receptor.
母鸡输卵管的孕酮受体由两个不同的激素结合多肽亚基组成，即A亚基（分子量为79,000）和B亚基（分子量为108,000）。我们使用高度纯化的B亚基制剂在体外免疫小鼠脾细胞。培养5天后，将这些细胞与SP2/0-Ag 14骨髓瘤细胞融合。使用酶联免疫吸附固相测定法筛选所得的杂交瘤，并通过有限稀释法克隆那些产生与免疫原结合抗体的杂交瘤。其中一个这样的克隆9B3-12分泌免疫球蛋白IgM类抗体，该抗体与B亚基结合。这通过抗体增加B亚基沉降系数速率的能力得以表明。此外，当B制剂中的蛋白质通过电泳分离并印迹到硝酸纤维素滤膜上时，该抗体与108,000道尔顿的蛋白质结合。9B3-12产生的抗体也与A亚基以及人孕酮受体发生反应，但在没有其受体的情况下不能与鸡肝糖皮质激素受体或孕酮结合。
