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US 7,083,911 B2Granted
Method for detection of ATP
AssigneePromega Corporation
InventorsKeith Wood, Rita Hannah, Richard A. Moravec
Life of this patent
Estimating the term…
Abstract
Methods and kits for detecting the presence of ATP, for measuring ATP concentrations, and for detecting viable cells using a composition comprising an ATP-dependent enzyme and one or more ATPase inhibitors.
Claims64
A method of detecting ATP in a sample comprising:
- (a) adding to the sample a reagent composition comprising one or more detergents and a luciferase,
- wherein the reagent composition is capable of maintaining at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents;
- and
- (b) detecting luminescence.
1 more element
A method of detecting ATP in a sample comprising:
- (a) adding to the sample a reagent composition comprising one or more detergents and a luciferase,
- wherein the reagent composition is capable of maintaining at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents;
- and
- (b) quantifying luminescence.
1 more element
A method of measuring cell viability within a population of cells comprising:
- (a) contacting the population of cells with a reagent composition comprising one or more detergents and a luciferase,
- wherein the reagent composition is capable of maintaining at least about 30% activity, as measured by luminescence after the reagent composition is combined with the population of cells, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the population of cells by at least about 25% relative to the population of cells' ATPase activity in the absence of the one or more detergents;
- and
- (b) detecting luminescence, wherein the amount of luminescence detected is proportional to the viability of the cells within the population.
1 more element
A method of determining the effect of a compound on a first population of cells comprising:
- (a) contacting the first population of cells with a concentration of the compound;
- (b) subsequently contacting the first population of cells with a reagent composition comprising one or more detergents and a luciferase,
- wherein the composition is capable of maintaining at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents;
- and
- (c) detecting an amount of luminescence;
- and
- (d) comparing the amount of luminescence in the first population to an amount of luminescence in a second population of cells.
4 more elements
A method of detecting ATP in a sample comprising:
- (a) adding to the sample a reagent composition comprising one or more detergents and a luciferase, wherein the luciferase comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:
- 1, 2, 3, and 4,
- wherein the reagent composition maintains at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents:
- and
- (b) detecting luminescence.
2 more elements
A method of detecting ATP in a sample comprising:
- (a) adding to the sample a reagent composition comprising one or more detergents and a luciferase, wherein the reagent composition further comprises NaF,
- wherein the reagent composition maintains at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents;
- and
- (b) detecting luminescence.
1 more element
A method of detecting ATP in a sample comprising:
- (a) adding to the sample a reagent composition comprising one or more detergents and a luciferase, wherein the reagent composition further comprises NaF,
- wherein the reagent composition maintains at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents;
- and
- (b) quantifying luminescence.
1 more element
A method of measuring cell viability within a population of cells comprising:
- (a) contacting the population of cells With a reagent composition comprising one or more detergents and a luciferase, wherein the reagent composition further comprises NaF,
- wherein the reagent composition maintains at least about 30% activity, as measured by luminescence after the reagent composition is combined with the population of cells, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the population of cells by at least about 25% relative to the population of cells' ATPase activity in the absence of the one or more detergents;
- and
- (b) detecting luminescence, wherein the amount of luminescence detected is proportional to the viability of the cells within the population.
1 more element
A method of determining the effect of a compound on a first population of cells comprising:
- (a) contacting the first population of cells with a concentration of the compound;
- (b) subsequently contacting the first population of cells with a reagent composition comprising one or more detergents and a luciferase, wherein the reagent composition further comprises NaF,
- wherein the composition maintains at least about 30% activity, as measured by luminescence after the reagent composition is combined with the sample, for at least one hour compared to the reagent composition's activity just after the luciferase is combined with the one or more detergents, and wherein the one or more detergents present in the reagent composition are collectively able to reduce ATPase activity endogenous to the sample by at least about 25% relative to the sample's ATPase activity in the absence of the one or more detergents;
- and
- (c) detecting an amount of luminescence;
- and
- (d) comparing the amount of luminescence in the first population to an amount of luminescence in a second population of cells.
4 more elements
Dependencies are inferred from the claim text.