3-(p-hydroxyphenyl)propionic acid as a new fluorogenic reagent for amine oxidase assays.
A detailed procedure of a new and extremely sensitive fluorometric assay for amine oxidases is presented. Hydrogen peroxide, produced by the oxidase reaction, reacted with 3-(p-hydroxyphenyl)propionic acid in the presence of peroxidase to yield a fluorescent compound by which enzyme activity could be determined. The enzyme reaction was terminated by NaOH solution, which increased the fluorescence intensity three- to fivefold. The detection limit thus obtained was as little as 0.02 nmol. The alkalinization also contributed to stopping the enzyme reaction and to the clarification of assay mixtures containing turbid enzyme preparations.
D006631:Amine Oxidase (Copper-Containing) / Q000032:analysis; D000818:Animals; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007668:Kidney / Q000201:enzymology; D008297:Male; D008928:Mitochondria / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D008995:Monoamine Oxidase / Q000032:analysis; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000032:analysis*; D010544:Peroxidases; D010666:Phenylpropionates; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence
D007202:Indicators and Reagents; D010666:Phenylpropionates; C008869:phloretic acid; D006861:Hydrogen Peroxide; D010544:Peroxidases; D006631:Amine Oxidase (Copper-Containing); D008995:Monoamine Oxidase; D000587:Oxidoreductases Acting on CH-NH Group Donors