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A novel assay for determination of androgenic or anti-androgenic activity of a serum sample or a test compound

机译:一种测定血清样品或测试化合物的雄激素或抗雄激素活性的新方法

摘要

The present invention is directed to methods for determining the androgenic or anti-androgenic activity of a sample and to a cell useful in these methods. To determine androgenic activity, the sample is contacted and incubated with a cell comprisingi) a luciferase reporter plasmid,ii) a fusion protein comprising a ligand-binding domain of the androgen receptor and a Gal4 DNA-binding domain, said fusion protein being able to bind to binding sites in said luciferase reporter plasmid,iii) a fusion protein comprising an N-terminal region of the androgen receptor and a transcriptional activation domain, andiv) an androgen receptor-interacting protein 3.The cell is then lysed and the luciferase activity of the lysate is measured. The measured luciferase activity is compared to that obtained by repeating the above method except that a control is added instead of the sample, to give the relative luciferase activity of the sample. The relative luciferase activity is used to detect or quantify an active androgen in the sample.
机译:本发明涉及确定样品的雄激素或抗雄激素活性的方法以及在这些方法中有用的细胞。为了确定雄激素活性,将样品接触并与包含i)荧光素酶报道质粒,ii)包含雄激素受体的配体结合结构域和Gal4 DNA结合结构域的融合蛋白的细胞一起孵育,所述融合蛋白能够结合至所述荧光素酶报道质粒中的结合位点,iii)包含雄激素受体N端区域和转录激活域的融合蛋白,iv)与雄激素受体相互作用的蛋白3.然后裂解细胞并进行荧光素酶活性分析测量裂解物的量。将测得的萤光素酶活性与通过重复上述方法获得的萤光素酶活性进行比较,不同之处在于加入对照而不是样品,以得到样品的相对萤光素酶活性。相对荧光素酶活性用于检测或定量样品中的活性雄激素。

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