Abstract: A method and kit are provided for enhancing the tolerance of an assay reagent to compounds in an assay sample, the assay reagent including a luciferase enzyme. The method includes contacting the luciferase with a tolerance enhancement agent in an amount sufficient to substantially protect luciferase enzyme activity from interference of the compound and minimize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Abstract: A modified luciferase protein which is a sensor for molecules including cAMP is provided. The modified luciferase protein includes one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with cAMP.

Abstract: The invention relates to compositions and methods for suppressing the luminescence intensity decline that occurs as the reaction between Cypridina luciferase and luciferin proceeds.

Abstract: A method and kit are provided for enhancing the tolerance of an assay reagent to compounds in an assay sample, the assay reagent including a luciferase enzyme. The method includes contacting the luciferase with a tolerance enhancement agent in an amount sufficient to substantially protect luciferase enzyme activity from interference of the compound and minimize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Abstract: The invention relates to methods, reagents and kits for detecting enzyme activity using bioluminescence. In particular, it relates to a novel luciferase assay system with reduced background luminescence to allow for increased detection sensitivity. Provided is a method of detecting luciferase activity in a sample using coelenterazine or an analog thereof as a substrate, comprising: (a) initiating luciferase-catalyzed luminescence production by contacting said sample with a luciferase detection reagent to yield a reaction mixture, said reagent comprising coelenterazine and at least one iodide source in an amount sufficient to reduce the autoluminescence of said coelenterazine, (b) incubating said reagent mixture under conditions suitable to produce luminescence, and (c) measuring the luminescence produced. Also provided are detections reagents and kits for use in such a method.

Abstract: A fusion gene is provided comprising the cDNA of Renilla luciferase and the cDNA of the "humanized" Aequorea green fluorescent protein. The fusion gene was used to produce a novel protein, the "Renilla-GFP fusion protein," which displayed both the luciferase activity of Renilla luciferase, and the green fluorescence of GFP. The Renilla-GFP fusion gene is useful as a double marker for monitoring gene expression quantitatively in UV light and by enzyme activity.

Abstract: The present invention provides a polynucleotide or polynucleotides encoding Oplophorus luciferase which is composed of 19 kDa and 35 kDa proteins, or the 19 kDa photoprotein, the recombinant secretional Oplophorus luciferase or the 19 kDa photoprotein encoded by the polynucleotide(s), an expression vector containing the polynucleotide(s) and a host transformed with the vector. Further, the invention provides a method for producing the recombinant Oplophorus luciferase or the photoprotein. These proteins could be recombinantly produced by culturing the host cell or by in vitro translation system using the recombinant expression vector.

Abstract: The invention relates to methods and kits for detecting enzyme activity using bioluminescence. In particular, it relates to a novel assay system with increased light yield for a sensitive and convenient detection of luciferase activity, such as luciferase reporter enzyme activity. Provided is a method of detecting luciferase activity in a sample, comprising incubating the sample in the presence of luciferin and ATP to allow the generation of a light signal, wherein said light signal is enhanced by performing the incubation in a reaction mixture comprising phosphate and ammonium ions, and measuring the light signal. The invention also relates to kits for use in such method.

Abstract: The invention relates to methods, reagents and kits for detecting enzyme activity using bioluminescence. In particular, it relates to a novel luciferase assay system with reduced background luminescence to allow for increased detection sensitivity. Provided is a method of detecting luciferase activity in a sample using coelenterazine or an analog thereof as a substrate, comprising: (a) initiating luciferase-catalyzed luminescence production by contacting said sample with a luciferase detection reagent to yield a reaction mixture, said reagent comprising coelenterazine and at least one iodide source in an amount sufficient to reduce the autoluminescence of said coelenterazine, (b) incubating said reagent mixture under conditions suitable to produce luminescence, and (c) measuring the luminescence produced. Also provided are detections reagents and kits for use in such a method.

Abstract: The invention relates to methods and kits for detecting enzyme activity using bioluminescence. In particular, it relates to a novel assay system with increased light yield for a sensitive and convenient detection of luciferase activity, such as luciferase reporter enzyme activity. Provided is a method of detecting luciferase activity in a sample, comprising incubating the sample in the presence of luciferin and ATP to allow the generation of a light signal, wherein said light signal is enhanced by performing the incubation in a reaction mixture comprising phosphate and ammonium ions, and measuring the light signal. The invention also relates to kits for use in such method.

Abstract: Nucleic acid compositions and polypeptides encoding a red-shifted form of firefly luciferase are provided. These red-shifted luciferases are characterized by spectrum of light emission having detectable emissions at 610 nm (luc610), preferably a primary peak at 610 nm. The nucleic acid compositions find use in various systems as a reporter gene, and are of particular interest for use as a reporter with in vivo systems, because of the efficient transfer of red light through tissues. The red-shifted luciferase may be combined in such assays with luciferases emitting at other spectra, in order to monitor multiple processes simultaneously.

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.

Abstract: A modified beetle luciferase protein which is an environmentally sensitive reporter protein is provided.

Abstract: A method and kit is provided for enhancing the tolerance of an assay reagent to compounds in an assay sample, the assay reagent including a luciferase enzyme. The method includes contacting the luciferase with a tolerance enhancement agent in an amount sufficient to substantially protect luciferase enzyme activity from interference of the compound and minimize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Abstract: A modified beetle luciferase protein which is an environmentally sensitive reporter protein is provided.

Abstract: The present invention generally relates to a methods, compositions and assays for real-time monitoring of the progression of a disease, such as a cancer in a subject, by measuring the level of bioluminescence signal in a biological sample obtained from a subject, where the bioluminescence signal is from a secreted luciferase protein expressed by a cell or tissue in the subject. One aspect of the present invention relates to administering to a subject a nucleic acid encoding a secreted luciferase, and in some embodiments, a disease or a diseased tissue such as tumor cells expresses the secreted luciferase protein, which is monitored in a biological sample, such as blood or urine obtained from a subject. One aspect of the invention relates to analysis of a secreted luciferase protein by measuring the level in a biological sample obtained from the subject without the need for invasive monitoring procedures.

Abstract: The present invention relates to an isolated nucleic acid and polypeptide sequence that encodes for a luciferase of Luciola italica, as well as mutants thereof. The luciferase proteins of the present invention have been found to have extended bioluminescence emission that is red- or blue-shifted, and are useful as a bioluminescent marker or as an additive to selected materials.

Abstract: An object of the present invention is to produce a luminescent probe that has less biological effects, efficiently emits visible to near-infrared light, which is excellent for the imaging of individuals, and the use thereof. The present invention provides a sugar chain-containing-luciferase derivative, wherein an organic fluorescent dye is bonded to the luciferase through the sugar chain.

Abstract: The present invention relates to an isolated nucleic acid and polypeptide sequence that encodes for a luciferase of Luciola italica, as well as mutants thereof. The luciferase proteins of the present invention have been found to have extended bioluminescence emission that is red- or blue-shifted, and are useful as a bioluminescent marker or as an additive to selected materials.

Abstract: An assay for the presence of luciferase in a biological sample offers heightened sensitivity, signal intensity and persistence. The assay is sensitive down to 50 fg luciferase. The biological sample is combined with essential ingredients luciferin, ADP, myokinase and Mg++. The myokinase converts ADP to ATP, necessary for the luciferase reaction, and AMP, which retards the reaction kinetics. The resulting assay exhibits a persistent glow emission which makes it adaptable to automation.