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| | ER-beta Radioligand Binding Assay and ERE-Luciferase Reporter Assay. |
| Description: | Radioligand binding assay was performed by using 96-well microtiterplates containing ER, 17beta-estradiol, and the test compound to be tested and SPAbeads. After incubation at room temperature for 2-4 h, the reaction was terminated by centrifugation. The radioactivity was counted in a Packard Topcount scintillation counter. Nonspecific binding was defined as the remaining radioactivity in the presence of 10 uM nonradioactive 17beta-estradiol. Assays were performed in triplicate. HELNbeta, a human cervix adenocarcinoma cell line derived from HeLa cells stably transfected with the reporter gene and the expression plasmid ERbeta, were used to quantify the antiestrogenic and estrogenic effects of compounds on ERE. |
| Affinity data for this assay | |
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