Method for enhancing activity of luciferase with fluorescence activity
AssigneeChisso Corporation
InventorsSatoshi Inouye, Satoko Sasaki
Life of this patent
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Abstract
Methods for enhancing luminescence of a luciferase (BFP-aq) with fluorescence activity derived from a calcium-binding photoprotein are provided. To a luciferase solution with fluorescence activity that contains an apoprotein, a calcium-binding photoprotein, which is constituted such that a coelenteramide or an analog thereof is coordinated inside, a coelenterazine that is the luminescent substrate of the luciferase or an analog thereof and a compound (e.g., imidazole etc.) having the function of removing an —NH-proton of the pyrazine ring of the imidazopyrazine skeleton in the coelenterazine or the analog thereof are added.
Claims17
A method for enhancing luminescence of a luciferase with fluorescence activity that contains an apoprotein in which a coelenteramide or an analog thereof is coordinated, comprising adding to a solution of the luciferase with the fluorescence activity a coelenterazine that is a luminescent substrate of the luciferase or an analog thereof and a compound having the function of removing an —NH— proton of the pyrazine ring of the imidazopyrazine skeleton in the coelenterazine or the analog thereof.
Use of a compound having the function of removing an —NH— proton of the pyrazine ring of the imidazopyrazine skeleton in a coelenterazine or an analog thereof for preparing a luminescence-enhancing agent for enhancing light emission generated from a luciferase with a fluorescence activity that contains an apoprotein in which a coelenteramide or an analog thereof is coordinated.
A kit comprising:
- a luciferase with fluorescence activity that contains an apoprotein in which a coelenteramide or an analog thereof is coordinated;
- a coelenterazine that is the luminescent substrate of the luciferase or an analog thereof;
- and
- a luminescence-enhancing agent that can enhance light emission of the luciferase,
- wherein the luminescence-enhancing agent is a compound having the function of removing an —NH— proton of the pyrazine ring of the imidazopyrazine skeleton in the coelenterazine or the analog thereof.
A method for enhancing luminescence of a BFP-aq that contains an apoaequorin in which a coelenteramide is coordinated, comprising adding a hcp-coelenterazine and imidazole to a solution of the BFP-aq.
Dependencies are inferred from the claim text.