摘要:
This invention provides bioluminescence resonance energy transfer (BRET) system that comprises four parts: 1) a bioluminescent protein that has luciferase activity; 2) an acceptor fluorophore that can accept the energy from the bioluminescent protein when they are associated, in the presence of the appropriate substrate; 3) a modulator that influences the proximity or the orientation of the bioluminescent protein and the fluorophore, and 4) an appropriate substrate to activate the luciferase activity of the bioluminescent protein. The components of this system interact to influence the spatial relationship between the bioluminescent protein and the fluorophore, that is demonstrated by the light emission from the system. The modulator can be a single entity, covalently attached to both the bioluminescent protein and the fluorophore, it can be two separate entities, each linked covalently to either the bioluminescent protein or the fluorophore, or an alternative configuration that falls within the scope of the invention. Thissystem can be used in both in vivo or in vitro assays to detect molecular changes in a wide variety of applications, and is amenable to automation. In particular, it is useful for assaying protein interactions, enzyme activities and the concentration of analytes or signaling molecules in cells or in solution.
摘要:
Luminescence assays and compositions for assay of biomolecular interaction and activity and detection of modulators of biomolecular interaction and activity are provided. Technology described herein has utility in a variety of assay formats and types, for example, simultaneous monitoring multiple parameters which affect interaction and activity of biological molecules. Compositions and methods are provided herein which include a first solid-phase support associated with a first specific binding agent and a photosensitizer,; a second solid-phase support associated with a second specific binding agent and a first emission system; and a third solid- phase support associated with a third specific binding agent and a second emission system.
摘要:
Methods for determining the presence or absence of a plurality of analytes in a sample with dual-acceptor time-resolved fluorescence resonance energy transfer are provided.
摘要:
Methods for determining the presence or absence of a plurality of analytes in a sample with dual-acceptor time-resolved fluorescence resonance energy transfer are provided.
摘要:
The present invention provides a bioluminescence resonance energy transfer (BRET) detection system characterised by a broad spectral resolution between donor and acceptor emission wavelengths. The broad spectral resolution between the emission wavelength of the bioluminescent donor protein and the fluorescent acceptor molecule results in an increased signal-to-base ratio and dynamic range in comparison with a basic BRET system.