Lofgren J A, Schwall R, Schmelzer C, Wong W L
Department of Immunology Research & Assay Technology, Genentech Inc., South San Francisco, California 94080.
J Immunoassay. 1991;12(4):565-78. doi: 10.1080/01971529108053280.
A goat antiserum to purified recombinant human activin A (rhAct-A), a dimer formed by two beta A-subunits of inhibin, has been produced. The immunoreactivity of the antiserum has been evaluated in an antigen coated enzyme-linked immunosorbent assay, in a radioimmunoassay using iodinated rhAct-A, and by Western blot analysis. The antiserum demonstrated some cross reactivity to inhibin A, a structurally related heterodimer which contains an identical beta A-subunit coupled to a distinct, though similar, alpha subunit. A simple radioimmunoassay for rhAct-A in tissue culture supernatant has been developed with rhAct-A affinity column purified polyclonal antiserum. The assay is precise and sensitive with a range of 0.31-40 ng/ml. The cross reactivity of inhibin A in the RIA is about 4.3%. Despite its cross-reactivity this antiserum will facilitate studies of the physiology of activin A and inhibin A which includes a Western blot analysis where a molecular size distinction is accomplished.
已制备出针对纯化重组人激活素A（rhAct-A）的山羊抗血清，rhAct-A是由抑制素的两个βA亚基形成的二聚体。已通过包被抗原的酶联免疫吸附测定、使用碘化rhAct-A的放射免疫测定以及蛋白质免疫印迹分析评估了该抗血清的免疫反应性。该抗血清对抑制素A表现出一些交叉反应性，抑制素A是一种结构相关的异二聚体，它包含一个与不同但相似的α亚基偶联的相同βA亚基。利用rhAct-A亲和柱纯化的多克隆抗血清开发了一种用于检测组织培养上清液中rhAct-A的简单放射免疫测定法。该测定法精确且灵敏，检测范围为0.31 - 40 ng/ml。放射免疫测定中抑制素A的交叉反应性约为4.3%。尽管存在交叉反应性，但这种抗血清将有助于对激活素A和抑制素A的生理学研究，包括进行蛋白质免疫印迹分析以实现分子大小区分。
