Detecting molecular interactions by fluorescence resonance energy transfer on a solid-phase support

摘要

The present disclosure features a variety of compositions, kits, and methods that are useful for, inter alia, detecting and/or analyzing an interaction between two molecules, a target molecule and a target-specific binding agent which can be, e.g., proteins, nucleic acids, saccharides or polysaccharides, small molecules, or combinations of any of the foregoing. The compositions, kits, and methods can also be used, e.g., to detect the presence or absence of an enzymatic activity (e.g., a kinase activity, a protease activity, or a phosphatase activity) in a sample; to identify a compound that modulates an interaction between two molecules; or to identify compounds that modulate the activity of an enzyme.

主权项

1. A composition comprising:
a first solid-phase support comprising a target-selective binding agent and a first member of a Fluorescence Resonance Energy Transfer (FRET) pair; and a target molecule associated with a second member of the FRET pair, wherein the target molecule is not associated with a solid-phase support; wherein the first solid-phase support comprises phthalocyanine and thioxene; wherein the first member of the FRET pair is a lanthanide chelate; and wherein the target-selective binding agent comprises deferoxamine-Fe3+ and preferentially binds to a target molecule that comprises a phosphomonoester moiety as compared to a target molecule that does not comprise a phosphomonoester moiety.