title: A novel fluorometric assay for quantitative analysis of dihydrofolate reductase activity in biological samples.
abstract: In an effort to study the level of dihydrofolate reductase (DHFR), the main molecular target of antifolate drugs, in healthy and malignant tissues of human origin, a new and convenient fluorometric enzymatic assay has been developed. The technique measures the overall decrease in fluorescence emission at 454 nm (lambda ex = 342 nm) due to the contributions from coenzyme oxidation and substrate reduction. This technique was developed by using an enzyme purified from beef liver. All criteria of quality were checked: sensitivity, reproducibility and specificity made it suitable for low activity measurements. It was successfully applied to human tissue crude extracts.
authors: Laboratoire de Physique Pharmaceutique, Faculté de Pharmacie, Marseille, France.
authors: O
authors: O
authors: Rimet
authors: M
authors: M
authors: Chauvet
authors: M
authors: M
authors: Bourdeaux
authors: C
authors: C
authors: Briand
mesh_terms: D000818:Animals; D002417:Cattle; D007202:Indicators and Reagents; D008099:Liver / Q000201:enzymology*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism*
chemical_list: D007202:Indicators and Reagents; D013762:Tetrahydrofolate Dehydrogenase