초록 ▼

A new peroxidase assay for substances demonstrating peroxidative activity is described which is based on the peroxidase catalyzed oxidation of dichlorofluorescin to dichlorofluorescein by peroxide. The dichlorofluorescin is formed from diacetyldichlorofluorescin either by the direct addition of pero

A new peroxidase assay for substances demonstrating peroxidative activity is described which is based on the peroxidase catalyzed oxidation of dichlorofluorescin to dichlorofluorescein by peroxide. The dichlorofluorescin is formed from diacetyldichlorofluorescin either by the direct addition of peroxide or by the inclusion of a peroxide generating system (e.g., glucose plus glucose oxidase or uric acid and uricase) in the reaction mixture. As long as all reactants other than the peroxidase are present in excess, the rate of fluorescence increase is linearly related to the amount of peroxidase in the sample under assay.

대표청구항 ▼

A composition useful for the determination of a substance having peroxidative activity, said composition, consisting essentially of diacetyldichlorofluorescin and a source of hydrogen peroxide in a concentration of from about 10-6 to 10-2 M, the ratio of diacetyldichlorofluorescin to hydrogen peroxi

A composition useful for the determination of a substance having peroxidative activity, said composition, consisting essentially of diacetyldichlorofluorescin and a source of hydrogen peroxide in a concentration of from about 10-6 to 10-2 M, the ratio of diacetyldichlorofluorescin to hydrogen peroxide or generated hydrogen peroxide ranging from 1:5 to 1:100. A method for assaying peroxidase comprising the steps of (a) contacting a substantially dry element comprising a reagent having a composition consisting essentially of diacetyldichlorofluorescin and a dry source of hydrogen peroxide at a concentration of from about 10-6 to 10-2 M with a sample for analysis, the ratio of diacetyldichlorofluorescin to generated hydrogen peroxide ranging from 1:5 to 1:100; and (b) detecting the dichlorofluorescein resulting from said contact. A method of assaying for antigen or antibody present in a sample to be tested comprising: (a) labeling a known amount of antigen or antibody with peroxidase and adding said labeled antigen or antibody with said sample to be tested to a known amount of corresponding antigen or antibody; and (b) determining the amount of unknown antigen or antibody by (i) contacting in an aqueous medium the mixture of (a) and a composition consisting essentially of diacetyldichlorofluorescin and a source of hydrogen peroxide at a concentration of from about 10-6 to 10-2 M to produce dichlorofluorescein, the ratio of diacetyldichlorofluorescin to hydrogen peroxide or generated hydrogen peroxide ranging from 1:5 to 1:00; (ii) detecting the dichlorofluorescein.