A solid-phase competitive radioimmunoassay for the insulin receptor.
A solid-phase competitive radioimmunoassay for the insulin receptor.
复制标题
胰岛素受体的固相竞争性放射免疫测定。
DOI：
10.1016/0003-2697(86)90322-2
复制
发表时间：
1986
影响因子：
2.9
通讯作者：
Beaudoin,J
中科院分区：
文献类型：
--
作者：
Roth,RA;Beaudoin,J
胰岛素受体的放射免疫测定法已经发展起来。在该试验中，未标记受体与125 I标记受体竞争结合固定在包被亲和纯化抗小鼠免疫球蛋白G的微量威尔斯孔上的单克隆抗受体抗体。该测定具有高度的重现性，可检测7 ng（14 fmol）的胰岛素受体。通过利用针对受体不同抗原区域的单克隆抗体，可以检查受体分子的不同部分。通过利用针对受体胞质结构域的抗体，开发了一种不受胰岛素存在影响的测定法，该测定法可同等检测不同种属（大鼠和人）和不同组织（胎盘和脑）的胰岛素受体。通过利用针对受体的自磷酸化位点的抗体，该测定显示能够检测受体的磷酸化程度。最后，该测定用于监测胰岛素处理的人淋巴细胞裂解物中胰岛素受体的减少。这种放射免疫测定法对于在各种生理条件下监测胰岛素受体的数量和状态都是有用的。
A radioimmunoassay for the insulin receptor has been developed. In this assay, unlabeled receptor competes with125I-labeled receptor for binding to monoclonal anti-receptor antibodies immobilized on microtiter wells coated with affinity-purified anti-mouse immunoglobulin G. This assay was highly reproducible and could detect 7 ng (14 fmol) of insulin receptor. By utilizing monoclonal antibodies to various antigenic regions of the receptor, different parts of the receptor molecule could be examined. By utilizing antibodies to the cytoplasmic domain of the receptor, an assay was developed which was not influenced by the presence of insulin and could equally detect the insulin receptor from different species (rat and human) and different tissues (placenta and brain). By utilizing antibodies to an autophosphorylation site of the receptor, the assay was shown capable of detecting the extent of phosphorylation of the receptor. Finally, this assay was utilized to monitor the decrease in insulin receptors in lysates of insulin-treated human lymphocytes. This radioimmunoassay should be useful for monitoring both the number and status of the insulin receptor under a variety of physiological conditions.