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    • 1. 发明申请
    • GENERIC KINASE/PHOSPHATASE ASSAY WITH SINGLE READOUT
    • 一次性发条/ PHOSPHATASE测定与单一读取
    • WO2009033580A1
    • 2009-03-19
    • PCT/EP2008/007122
    • 2008-09-01
    • F. HOFFMANN-LA ROCHE AGENDERLE, ThiloROTH, Doris
    • ENDERLE, ThiloROTH, Doris
    • C12Q1/42C12Q1/48G01N33/542
    • G01N33/542C12Q1/42C12Q1/485
    • The present invention relates to a generic method for detecting a kinase or phosphatase activity. The method comprises the following steps: incubating a kinase or phosphatase activity sample with a kinase or phosphatase substrate molecule comprising either a fluorophore having a detectable readout or molecule with an aromatic group which serves as a quencher of the fluorophore, incubating the mixture of step a) with a detection entity comprising either a fluorophore or a molecule with an aromatic group and a binding partner, wherein the substrate molecule and the detection entity are capable of binding to the binding partner and the binding of the substrate molecule and the detection entity to the binding partner lead to an altered readout of the fluorophore, and measuring the readout of the fluorophore in the mixture of step b), wherein an altered readout of the fluorophore compared to a blank is indicative for the presence of a kinase or phosphatase activity in the sample.
    • 本发明涉及用于检测激酶或磷酸酶活性的通用方法。 该方法包括以下步骤:将激酶或磷酸酶活性样品与激酶或磷酸酶底物分子孵育,所述激酶或磷酸酶底物分子包含具有可检测的读出的分子或具有用作荧光团的猝灭剂的芳族基团的分子,孵育步骤a的混合物 ),其具有包含荧光团或具有芳族基团的分子和结合配偶体的检测实体,其中所述底物分子和所述检测实体能够结合所述结合配偶体,并且所述底物分子与所述检测实体与所述检测实体的结合 结合配偶体导致荧光团的改变的读数,以及测量步骤b)的混合物中的荧光团的读数,其中与空白相比,荧光团的改变的读数指示在所述荧光团中存在激酶或磷酸酶活性 样品。