Hoffmann-la Roche Inc.

F. Hoffmann-la Roche Ag

Doris Roth

Thilo Enderle

Hugues Matile

The present invention relates to an in vitro method for detecting cAMP or cGMP comprising a) contacting a mixture with a complex of a tracer and a dequencher, wherein the tracer is a fluorophore covalently linked to a cAMP quencher, and b) measuring the change in fluorescence. Furthermore, the present invention relates to the use of said method for determining the cAMP or cGMP concentration in the mixture, for determining the activity of a receptor wherein the signal transduction of the receptor comprises cAMP or cGMP and for screening of a ligand for such a receptor.

Method of detection of cAMP and cGMP via a fluorophore and dequencher complex

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.

Generic kinase/phosphatase assay with single readout

The present invention relates to a method of determining kinase or phosphatase activity based on a three parts system. The method comprises contacting a binding partner which can bind phosphorylated peptides, a detection molecule and a substrate peptide. Determination of activities is achieved by measuring energy transfer between an energy donor and an energy acceptor that are present on the detection molecule and the substrate molecule.

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Basel, Switzerland

Doris Roth