| Assay Method Information | |
| | Coactivation of TR-FRET Farnesoid X Receptor Protein |
| Description: | Purchasing Invitrogen PV4833 Kit.Step 1, the compound was weighed and dissolved in 100% DMSO, the highest concentration is 3000 μM, and then diluted with DMSO by 3-fold serial dilution to get 10 concentrations.Step 2, the above formulated compound solutions were diluted 100 times with the buffer provided in the kit, which were mixed respectively and 10 μL of which were added into a 384 well plate in turn.Step 3, FXR recombinant nucleo receptor protein was diluted with a buffer to formulate 4× concentration, and 5 μL of which was added into the above 384 well plate containing the compound.Step 4, Fluorescein-SRC2-2 and Tb anti-GST antibody were diluted with a buffer respectively, both concentration are 4×, after the two reagents were mixed together, 10 μL of which was added into the 384 well plate described in step 3.At last, the solution in the 384 well plate was blend by centrifugation, and then incubated at room temperature for 1 hour. And then the detection was performed at 520, 495 and 337 nm wavelengths by TR-FRET endpoint method, ECso values were calculated using the detection values of ER=520 nm/495 nm wavelength. |
| Affinity data for this assay | |
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