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Miniaturization of a Panel of High Throughput Yeast-Cell-Based Nuclear Receptor Assays in 384- and 1536-Well Microplates

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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.

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Keywords: 17-estradiol; AR assay; AhR assays; BMAAREluc/AR; BMAAhR/ ARNT; BMAER; Bentzo-a-pyrene; Biomek NXP; Cell-based assay; D-luciferin; Dihydro- testosterone; ER yeast assay; GraphPad Prism 4; LOD; REACH; Victor3 1420 Multilabel Counter; Z' factor; endocrine-disrupting compounds; estrogen receptor; high throughput assay; high throughput screening; luciferase; nuclear receptors; saccharomyces cerevisiae

Document Type: Research Article

Publication date: 01 January 2011

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  • Combinatorial Chemistry & High Throughput Screening publishes full length original research articles and reviews describing various topics in combinatorial chemistry (e.g. small molecules, peptide, nucleic acid or phage display libraries) and/or high throughput screening (e.g. developmental, practical or theoretical). Ancillary subjects of key importance, such as robotics and informatics, will also be covered by the journal. In these respective subject areas, Combinatorial Chemistry & High Throughput Screening is intended to function as the most comprehensive and up-to-date medium available. The journal should be of value to individuals engaged in the process of drug discoveryand development, in the settings of industry, academia or government.
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