Method of permanent fluorescent assay
AssigneesNEN Life Science Products, Inc., Washington University
InventorsMark Norman Bobrow, Kevin Aaron Roth
Life of this patent
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Abstract
A method for performing a peroxidase-based assay by reacting a peroxidase enzyme with a soluble organic p-hydroxyphenyl-containing compound having the formula: wherein Q is a linear or branched 1-12 heteroatom alkyl wherein the heteroatoms are selected from C, N, O, and S, wherein the bonds connecting the heteroatom alkyl chain are single or double, wherein any carbon atom in the heteroatom alkyl chain optionally includes a substituent selected from —OH, —COOH, —NH2, and —SH, and wherein R is selected from —OH, —COOH, —NH2, and —CH3; and converting the soluble organic p-hydroxyphenyl-containing compound into a stable insoluble product which becomes highly fluorescent after illumination, upon excitation.
Claims13
A method for performing a peroxidase-based assay comprising:
- reacting a peroxidase enzyme with a soluble p-hydroxyphenyl-containing substrate compound selected from the group consisting of:
- said substrate compound being further characterized in that after reaction with said peroxidase enzyme it forms an insoluble product which is characterized in that when it is illuminated it is converted to an activated form;
- converting said insoluble product to an activated form by illuminating it with light;
- illuminating said activated form of said insoluble product with light of a first wavelength so that said activated product fluoresces at a second wavelength which is longer than said first wavelength;
- and
- detecting said second wavelength.
A peroxidase-based assay method for detecting the presence or absence of a desired substance in a sample, said method comprising the steps of:
- reacting a peroxidase enzyme with a soluble p-hydroxyphenyl-containing substrate, said substrate compound being selected from the group consisting of:
- said substrate compound being further characterized in that after reaction with said peroxidase enzyme it forms an insoluble product which is characterized in that when it is illuminated it is converted to an activated form;
- converting said stable, insoluble product to an activated form by illuminating it with light;
- illuminating said activated form of said insoluble product with light of a first wavelength so that said activated product fluoresces at a second wavelength which is longer than said first wavelength;
- and
- detecting said second wavelength.
In a peroxidase-based assay of the type wherein a peroxidase enzyme is reacted with a substrate, the improvement comprising in combination:
- said substrate being a p-hydroxyphenyl-containing substrate selected from the group consisting of:
Dependencies are inferred from the claim text.