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| | HTRF Coactivator Recruitment Assay |
| Description: | To each well of a 96-well assay plate, assay buffer was added containing purified GST-FXR, Eu-labeled anti-GST antibody (Perkin-Elmer), streptavidin-XL665 (Packard Instruments), biotin-tagged SRC-1 fragment. The binding reaction was initiated by the addition of compound. The plate fluorescence was read in a Discovery instrument (Packard) using an excitation wavelength of 337 nm and monitoring emission wavelengths of 620 and 665 nm. Data was expressed as the ratio of emission at 665/620 nm, were normalized with control wells to which 1 ul of DMSO was added instead of compound, and percentage maximal activity was calculated relative to controls with a saturating concentration (150 uM) of chenodeoxycholic acid (CDCA, Sigma-Aldrich). |
| Affinity data for this assay | |
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