Quenching reagents and assays for enzyme-mediated luminescence
AssigneePromega Corporation
InventorsBruce A. Sherf, Keith V. Wood, Elaine T. Schenborn
Life of this patent
Estimating the term…
Abstract
The present invention relates to single and dual-reporter luminescence assays utilizing general and specific reagents to quench enzyme-mediated reactions. In one embodiment of the invention, a reagent is added to the assay which non-specifically quenches enzyme-mediated luminescent reactions. In another embodiment of the invention, a reagent is added to the assay which simultaneously quenches one enzyme-mediated luminescent reaction while activating another distinct enzyme-mediated luminescent reaction. An assay kit containing specific quench reagents, and the reagents themselves are also disclosed.
Claims60
A method of assaying enzyme-mediated luminescence reactions comprising:
- (a) initiating at least one enzyme-mediated luminescence reaction;
- then
- (b) quantifying luminescence energy produced by the luminescence reaction;
- and then
- (c) quenching photon emission from the enzyme-mediated luminescence reaction by introducing at least one quench reagent to the luminescence reaction.
A method of assaying enzyme-mediated luminescence reactions within multisample assay plates comprising:
- (a) initiating at least one enzyme-mediated luminescence reaction;
- (b) quantifying luminescence energy produced by the luminescence reaction;
- and
- (c) quenching the enzyme-mediated luminescence reaction by introducing at least one quench reagent to the luminescence reaction;
- whereby refractive interference of light between samples within the multisample assay plate is eliminated.
An enzyme-mediated luminescence reaction assay kit comprising:
- at least one quench reagent composition capable of quenching photon emission from an enzyme-mediated luminescence reaction by a factor of at least 1,000-fold;
- a suitable first container, said at least one quench reagent composition disposed therein;
- at least one functional enzyme substrate corresponding to the enzyme-mediated luminescence reaction being assayed;
- a suitable second container, said at least one functional enzyme substrate disposed therein;
- and
- instructions for use.
A dual-reporter enzyme-mediated luminescence reaction assay kit comprising:
- a first functional enzyme substrate corresponding to a first enzyme-mediated luminescence reaction being assayed;
- a suitable first container, said first functional enzyme substrate disposed therein;
- a quench-and-activate composition comprising quench reagents capable of quenching photon emission from the first enzyme-mediated luminescence reaction by a factor of at least 1,000-fold, and a second and distinct functional enzyme substrate corresponding to a second and distinct enzyme-mediated luminescence reaction;
- a suitable:
- second container, said quench-and-activate composition disposed therein;
- and
- instructions for use.
Enzyme-mediated luminescence reaction quench reagents capable of reducing photon emissions from enzyme-mediated luminescence reactions comprising:
- iodide,
- iodine,
- sulfate,
- nitrate,
- iso-propanol,
- 2-(4-aminophenyl)-6-methylbenzothiazole,
- dimethyldecylphosphine oxide,
- pyrophosphate,
- benzothiazole,
- 2-phenylbenzothiazole,
- n-butanol,
- trans-1,2,-diaminocyclohexane-N,N,N',N'-tetraacetic acid,
- 2-(6'-hydroxy-2'-benzothiazolyl)-thiazole-4-carboxylic acid,
- ethylenediaminetetraacetic acid,
- 2(o-hydroxyphenyl)benzothiazole,
- India fountain pen ink,
- adenosine 5'-triphosphate 2',3'-acyclic dialcohol, periodate oxidized, borohydride reduced
- sodium dodecyl sulfate,
- citric acid,
- Tween.RTM.20,
- Triton.RTM.X-100,
- and mixtures thereof.
Dependencies are inferred from the claim text.