Method for enhancing detection of superoxide anion
AssigneeTrustees of Dartmouth College
InventorLorraine C. Pfefferkorn
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Abstract
A method for enhancing luminol chemiluminescence assays for superoxide anion comprising an orthovanadate salt, such as sodium orthovanadate. Orthovanadate enhances photon emission when luminol is cleaved by superoxide anion. The orthovanadate salt is non-cytotoxic and non-denaturing to intact cells. A method comprises reacting luminol in the presence of an orthovanadate anion with an analyte suspected of containing superoxide anion and measuring the photon output. A kit comprises a predetermined amount of luminol and orthovanadate in separate containers.
Claims16
A method for performing an enhanced chemiluminescence assay of superoxide anion, comprising:
- (a) providing an analyte suspected of containing or of being able to generate superoxide anion;
- (b) providing a predetermined amount of a chemiluminescent substance cleavable by superoxide anion to produce a signal;
- (c) providing a predetermined amount of orthovanadate anion;
- (d) contacting the analyte with the chemiluminescent substance in (b) in the presence of the orthovanadate anion in such a manner as to permit reaction of any superoxide anion present in the analyte with the chemiluminescent substance in (b) resulting in the chemical cleavage of the chemiluminescent substance in (b) and the release of photons under conditions that are non-denaturing and non-cytotoxic to intact cells;
- and
- (e) measuring the photons released in step (d).
A kit for performing an enhanced chemiluminescence assay of superoxide anion, comprising the following components in separate containers:
- (a) a predetermined amount of orthovanadate anion has been inserted after "comprising";
- in line 3 under conditions that are non-denaturing and non-cytotoxic to intact cells;
- and
- (b) a predetermined amount of a chemiluminescent substance cleavable by superoxide anion to produce a signal.
Dependencies are inferred from the claim text.