Miniaturization of a panel of high-throughput yeast-cell-based nuclear receptor assays in 384- and 1536-well microplates.
A panel of luminescent Saccharomyces cerevisiae cell-based nuclear receptor assays, consisting of human estrogen receptors α and β, androgen receptor, and aryl hydrocarbon receptor, was miniaturized from the standard 96-well microplate format to high-throughput 384- and 1536-well microplate formats. In these assays, firefly luciferase lacking the peroxisome targeting sequence was used as a reporter and D-luciferin substrate was pre-mixed with the yeast cells before the incubation step, eliminating cell lysis and substrate addition steps, and allowing multiple readings at any desired time point. All of the assays were highly functional in the 384-well format, and most functioned well in the 1536-well format. The detection limit of the estrogen receptor α assay was even lower in the miniaturized microplate formats than in the original 96-well format. The panel of yeast-cell-based nuclear receptor assays can be used for high-throughput chemical testing and environmental monitoring of potential endocrine-disrupting activity of compounds and samples.
Department of Food and Environmental Sciences, Faculty of Agriculture and Forestry, P.O. Box 56 (Viikinkaari 9), 00014 University of Helsinki (00790 Helsinki), Finland. johanna.rajasarkka@helsinki.fi
Johanna
J
Rajasärkkä
Marko
M
Virta
D017930:Genes, Reporter; D006801:Humans; D008904:Miniaturization; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D018160:Receptors, Cytoplasmic and Nuclear