Monoclonal antibody to chicken oviduct progesterone receptor.
Monoclonal antibody to chicken oviduct progesterone receptor.
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鸡输卵管孕酮受体单克隆抗体。
DOI：
10.1073/pnas.80.10.2854
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发表时间：
1983
影响因子：
11.1
通讯作者：
E. Baulieu
中科院分区：
文献类型：
--
作者：
C. Radanyi;I. Joab;J. Renoir;H. Richard;E. Baulieu
在Wistar大鼠体内培养钼酸盐稳定鸡输卵管孕酮受体抗体，并通过与均相放射性碘化孕酮受体的相互作用检测抗体。将该大鼠脾细胞与小鼠Sp2/0-Ag14骨髓瘤细胞融合，用125i标记受体双免疫沉淀法检测杂交细胞产生的抗体。然后通过限制稀释克隆一个阳性培养的细胞，并研究了一个杂交瘤细胞系。该单克隆抗体为IgG2b，它与钼酸盐稳定的“8-9S”形式的鸡输卵管孕酮受体反应，标记为[3H]孕酮或[3H]ORG 2058（一种高亲和力的合成孕激素）。8-9S孕酮受体的Kd约等于1 nM。孕酮受体-单克隆抗体复合物用放射性孕酮标记，提示抗体不阻止激素结合。通过使用[35S]蛋氨酸标记的抗体，我们能够独立检测黄体酮受体结合放射性激素的特征功能。与人黄体酮受体无交叉反应。
Antibodies to molybdate-stabilized chicken oviduct progesterone receptor were raised in a Wistar rat and detected by interaction with homogeneous radioiodinated progesterone receptor. Spleen cells of this rat were then fused with mouse Sp2/0-Ag14 myeloma cells and the antibodies produced by the hybrid cells were detected by double immunoprecipitation using the 125I-labeled receptor. Cells of one of the positive cultures were then cloned by limiting dilution and one hybridoma cell line was studied. The monoclonal antibody produced was an IgG2b, and it reacted with the molybdate-stabilized "8-9S" form of the chicken oviduct progesterone receptor, labeled with either [3H]progesterone or [3H]ORG 2058 (a high-affinity synthetic progestin). Kd for the 8-9S progesterone receptor was approximately equal to 1 nM. Progesterone receptor-monoclonal antibody complexes were labeled with radioactive progesterone, suggesting that antibody does not prevent hormone binding. By using a [35S]methionine-labeled antibody, we were able to detect the progesterone receptor independently of its characteristic function of binding radioactive hormone. No crossreaction with human progesterone receptor was detected.