Fluorescein-cyanine 5 as a fluorescence resonance energy transfer pair
First Claim
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1. A method for detecting the proximity of a first molecular segment to a second molecular segment, comprising:
- a) covalently attaching fluorescein to a first molecular segment;
b) covalently attaching cyanine 5 to a second molecular segment; and
c) detecting the presence or absence of fluorescein-induced emission of cyanine 5 as a result of fluorescence resonance energy transfer when the first molecular segment and second molecular segment are in proximity to each other.
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Abstract
A method is provided for detecting the proximity of a first molecular segment to a second molecular segment. The method comprises covalently attaching fluorescein to a first molecular segment, covalently attaching cyanine 5 to a second molecular segment, and detecting the presence or absence of fluorescein-induced emission of cyanine 5 as a result of fluorescence resonance energy transfer when the first molecular segment and second molecular segment are in proximity to each other. Compositions and compounds comprising the fluorescein and cyanine 5 pair are also provided.
46 Citations
44 Claims
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1. A method for detecting the proximity of a first molecular segment to a second molecular segment, comprising:
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a) covalently attaching fluorescein to a first molecular segment;
b) covalently attaching cyanine 5 to a second molecular segment; and
c) detecting the presence or absence of fluorescein-induced emission of cyanine 5 as a result of fluorescence resonance energy transfer when the first molecular segment and second molecular segment are in proximity to each other. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 39, 40, 41, 42, 43)
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26. A composition comprising a first member of a binding pair directly or indirectly attached to fluorescein and a second member of the binding pair directly or indirectly attached to cyanine 5, wherein the first and second members of the binding pair are associated so that the fluorescein and cyanine 5 are in fluorescence resonance energy transfer proximity to each other.
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38. A compound comprising a first molecular segment covalently bound to fluorescein and a second molecular segment covalently bound to cyanine 5.
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44. In a FRET-based assay having a dye pair, the improvement comprising using fluorescein and cyanine 5 as the dye pair.
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