Energy transfer assay method and reagent

Abstract

Disclosed is a non-fluorescent cyanine dye that may be used as an acceptor in fluorescence energy transfer assays involving the detection of binding and/or cleavage events in reactions involving biological molecules, and assay methods utilising such dyes. The non-fluorescent cyanine dye is a compound of formula (I), wherein the linker group Q contains at least one double bond and forms a conjugated system with the rings containing X and Y; groups R , R , R and R are attached to the rings containing X and Y, or optionally, are attached to atoms of the Z and Z ring structures; Z and Z each represent a bond or the atoms necessary to complete one or two fused aromatic rings each ring having five or six atoms, selected from carbon atoms and, optionally, no more than two oxygen, nitrogen and sulphur atoms; at least one of groups R , R , R , R , R , R and R is a target bonding group; any remaining groups R , R , R , R and R groups are independently selected from the group consisting of hydrogen, C -C alkyl, OR , COOR , nitro, amino, acylamino, quaternary ammonium, phosphate sulphonate and sulphate, where R is selected from H and C -C alkyl; any remaining R and R are selected from C -C alkyl which may be unsubstituted or substituted with phenyl, the phenyl being optionally substituted by up to two substituents selected from carboxyl, sulphonate and nitro groups; characterised in that at least one of the groups R , R , R , R , R , R and R comprises a substituent which reduces the fluorescence emission of said dye such that it is essentially non-fluorescent.