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• W2092072244 abstract "A new bioluminescence resonance energy transfer (BRET) homogeneous assay to evaluate the presence of estrogen-like compounds has been developed and optimized. The assay is based on the direct evaluation of estrogen α receptor (ERα) homodimerization as a result of estrogen-like compound binding. ERα monomer was genetically fused either to Renilla luciferase (Rluc) or to enhanced yellow fluorescent protein (EYFP). In the presence of estrogens, ERα dimerization brings Rluc and EYFP molecules close enough for an energy transfer. An in vitro BRET assay was first developed using purified fusion proteins (ERα-Rluc and ERα-EYFP) expressed in Escherichia coli to evaluate and optimize the analytical performances of the assay in the presence of 17-β estradiol. The “in vivo” BRET quantitative assay was then developed by coexpressing the two fusion proteins in live HepG2 cells. The assay can be performed in 96-well microplate format with a 30-min incubation and allows detection with adequate accuracy and precision of as low as 1 nM of 17-β estradiol. This new “in vivo” BRET assay allows evaluating the estrogen-like activity and synthetic xenoestrogens from biological and environmental samples." @default.
• W2092072244 created "2016-06-24" @default.
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• W2092072244 date "2004-10-26" @default.
• W2092072244 modified "2023-10-10" @default.
• W2092072244 title "Development of a Bioluminescence Resonance Energy-Transfer Assay for Estrogen-Like Compound in Vivo Monitoring" @default.
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• W2092072244 doi "https://doi.org/10.1021/ac048914h" @default.
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• W2092072244 hasPublicationYear "2004" @default.
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