Patent ID: 8309310
Filing Date: 2012-11-13
Classification: C12Q,G01N

Abstract:
1. A method for detecting a kinase activity or phosphatase activity comprising the steps: a) incubating a kinase or phosphatase activity sample with a kinase or phosphatase substrate molecule comprising a fluorophore having a detectable readout, b) incubating the mixture of step a) with a detection entity, comprising an aromatic group, and a binding partner, wherein the phosphorylated 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 wherein the binding of the substrate molecule and the detection entity to the binding partner involves ionic interactions, and c) 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, wherein the fluorophore is selected from the group consisting of Fluorescein, Rhodamine B, TMR, ATTO 590, ATTO 655, ATTO 680, Atto 700, MR 121, Bodipy 630/650 and Bodipy FL; and further wherein the aromatic group is selected from the group consisting of the amino acids tryptophan, tyrosine and phenylalanine.