Bioluminescent assay of femtomole levels of estrone and estradiol.
A rapid and very sensitive enzymatic assay of estrogens was developed using the transhydrogenase reaction of the human estradiol-17 beta dehydrogenase and bioluminescence for the quantification of NADH accumulated during the first reaction. The assay requires two steps: first, the addition of transhydrogenase buffer, and a few minutes later the addition of the bioluminescence reagent. The method can determine 0.1 to 50 pg of estrogens in the assay, in 1 h, with a coefficient of variation of 5%.
D015067:17-Hydroxysteroid Dehydrogenases; D002021:Buffers; D004958:Estradiol / Q000032:analysis* / Q000097:blood / Q000652:urine; D004960:Estradiol Dehydrogenases; D004970:Estrone / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D008163:Luminescent Measurements; D008832:Microchemistry; D009243:NAD / Q000032:analysis; D009247:NADH, NADPH Oxidoreductases; D009250:NADP Transhydrogenases
D002021:Buffers; D009243:NAD; D004970:Estrone; D004958:Estradiol; D015067:17-Hydroxysteroid Dehydrogenases; D004960:Estradiol Dehydrogenases; D009247:NADH, NADPH Oxidoreductases; D009250:NADP Transhydrogenases