The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.
The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.
Patent No.:
Date of Patent:
Apr. 07, 2015
Filed:
Jul. 30, 2009
Jurriaan Zwier, Rochefort du Gard, FR;
Robert Poole, Ringwood Hampshire, GB;
Herve Ansanay, Montpellier, FR;
Michel Fink, Bagnols sur Ceze, FR;
Eric Trinquet, Pont Saint Esprit, FR;
Jurriaan Zwier, Rochefort du Gard, FR;
Robert Poole, Ringwood Hampshire, GB;
Herve Ansanay, Montpellier, FR;
Michel Fink, Bagnols sur Ceze, FR;
Eric Trinquet, Pont Saint Esprit, FR;
Cis-Bio International, Gif sur Yvette Cedex, FR;
Abstract
The instant invention provides for methods for detecting the internalization of a transmembrane protein of interest expressed at the surface of a cell. More specifically, the methods involve (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 (1) a modulating agent which is a fluorescent FRET acceptor compound compatible with the fluorescent metal complex, the final concentration of which in the reaction medium is greater than 10M; or (2) a reducing agent, the redox potential of which is less than +0.1 V and preferably between 0.25 and 0.75 V; or (3) an agent which binds specifically, by non-covalent bonding, with the fluorescent metal complex; (d) adding a metal ion which competes with the rare earth so as to form a non-fluorescent 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 the compound is a fluorescent acceptor compound; and (e) comparing the signal measured in step d) with a reference signal measured on cells having been subjected only to steps a) and c).