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    Detecting cell membrane protein endocytosis
    5.
    发明授权
    Detecting cell membrane protein endocytosis 有权
    检测细胞膜蛋白内吞

    公开(公告)号：US07202043B2

    公开(公告)日：2007-04-10

    申请号：US11244256

    申请日：2005-10-04

    申请人： Yun-Jung Choi ,  Kun Peng ,  Linda Kauffman ,  Rajendra Singh

    发明人： Yun-Jung Choi ,  Kun Peng ,  Linda Kauffman ,  Rajendra Singh

    IPC分类号： G01N33/53

    CPC分类号： C12Q1/37 ,  G01N33/5005 ,  G01N33/567 ,  G01N2333/924 ,  G01N2333/976 ,  Y10S435/962 ,  Y10S435/963

    摘要： Methods and reagents are provided for determining endocytosis, using a cell expressing an externally polypeptide labeled cell membrane receptor, and as a reagent an antibody to said label conjugated to a fragment of an enzyme fragment complementation pair. Compounds are tested for their effect on endocytosis by complexing the reagent with said cells, adding the test compound at any time in relation to the complexing, allowing any endocytosis to occur, proteolytically degrading external enzyme fragment, adding protease inhibitor and the complementary member of the enzyme fragment complementation pair and substrate. The product provides a detectable signal related to the amount of endocytosis that occurred. The method is readily automated as all steps can occur in a single vessel without separations and washings.

    摘要翻译： 提供了用于确定内吞作用的方法和试剂，使用表达外部多肽标记的细胞膜受体的细胞，以及作为与所述酶标记片段互补对的片段缀合的所述标记的抗体作为试剂。 通过使试剂与所述细胞络合来测试化合物对内吞作用的影响，在任何时间加入测试化合物与复合相关，允许发生任何内吞作用，蛋白水解降解外部酶片段，加入蛋白酶抑制剂和补体成员 酶片段互补对和底物。 该产品提供与发生的胞吞作用量相关的可检测信号。 该方法易于自动化，因为所有步骤都可以发生在没有分离和洗涤的单个容器中。

    Detecting cell membrane protein endocytosis
    6.
    发明申请
    Detecting cell membrane protein endocytosis 有权
    检测细胞膜蛋白内吞

    公开(公告)号：US20060073523A1

    公开(公告)日：2006-04-06

    申请号：US11244256

    申请日：2005-10-04

    申请人： Yun-Jung Choi ,  Kun Peng ,  Linda Kauffman ,  Rajendra Singh

    发明人： Yun-Jung Choi ,  Kun Peng ,  Linda Kauffman ,  Rajendra Singh

    IPC分类号： G01N33/567 ,  G01N33/53

    CPC分类号： C12Q1/37 ,  G01N33/5005 ,  G01N33/567 ,  G01N2333/924 ,  G01N2333/976 ,  Y10S435/962 ,  Y10S435/963

    摘要： Methods and reagents are provided for determining endocytosis, using a cell expressing an externally polypeptide labeled cell membrane receptor, and as a reagent an antibody to said label conjugated to a fragment of an enzyme fragment complementation pair. Compounds are tested for their effect on endocytosis by complexing the reagent with said cells, adding the test compound at any time in relation to the complexing, allowing any endocytosis to occur, proteolytically degrading external enzyme fragment, adding protease inhibitor and the complementary member of the enzyme fragment complementation pair and substrate. The product provides a detectable signal related to the amount of endocytosis that occurred. The method is readily automated as all steps can occur in a single vessel without separations and washings.

    摘要翻译： 提供了用于确定内吞作用的方法和试剂，使用表达外部多肽标记的细胞膜受体的细胞，以及作为与所述酶标记片段互补对的片段缀合的所述标记的抗体作为试剂。 通过使试剂与所述细胞络合来测试化合物对内吞作用的影响，在任何时间加入测试化合物与复合相关，允许发生任何内吞作用，蛋白水解降解外部酶片段，加入蛋白酶抑制剂和补体成员 酶片段互补对和底物。 该产品提供与发生的胞吞作用量相关的可检测信号。 该方法易于自动化，因为所有步骤都可以发生在没有分离和洗涤的单个容器中。