发明申请
- 专利标题: METHOD FOR DETECTING MEMBRANE PROTEIN INTERNALIZATION
- 专利标题(中): 检测膜蛋白内含物的方法
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申请号: US13056738申请日: 2009-07-30
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公开(公告)号: US20120009599A1公开(公告)日: 2012-01-12
- 发明人: Jurriaan Zwier , Robert Poole , Herve Ansanay , Michel Fink , Eric Trinquet
- 申请人: Jurriaan Zwier , Robert Poole , Herve Ansanay , Michel Fink , Eric Trinquet
- 申请人地址: FR GIF SUR YVETTE CEDEX
- 专利权人: CIS-BIO INTERNATIONAL
- 当前专利权人: CIS-BIO INTERNATIONAL
- 当前专利权人地址: FR GIF SUR YVETTE CEDEX
- 优先权: FR0855296 20080731
- 国际申请: PCT/FR09/51538 WO 20090730
- 主分类号: G01N21/64
- IPC分类号: G01N21/64
摘要:
A method for detecting the internalization of a transmembrane protein of interest expressed at the surface of a cell, comprising the following steps: a) labelling the protein of interest with a fluorescent metal complex, the lifetime of which is greater than 0.1 ms; b) adding to the reaction medium a composition capable of causing the internalization of the protein of interest; c) adding to the reaction medium a modulating agent selected from: a. a fluorescent or nonfluorescent FRET acceptor compound compatible with said fluorescent metal complex, the final concentration of which in the reaction medium is greater than 10−7M; b. a reducing agent, the redox potential of which is less than +0.1 V and preferably between 0.25 and 0.75 V; c. an agent which binds specifically, by noncovalent bonding, with the fluorescent metal complex; d. a metal ion which competes with the rare earth so as to form a nonfluorescent metal complex; d) measuring the luminescence emitted by the reaction medium at the emission wavelength of the fluorescent metal complex and/or at the emission wavelength of the modulating compound when said compound is a fluorescent acceptor compound; e) comparing the signal measured in step d) with a reference signal measured on cells having been subjected only to steps a) and c).
公开/授权文献
- US08999653B2 Method for detecting membrane protein internalization 公开/授权日:2015-04-07
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