| Assay Method Information | |
| | Binding Activity of Compound to Progesterone Receptor PR |
| Description: | The binding activity of the compound to PR was determined using the LanthaScreen TR-FRET Progesterone Receptor Coactivator Assay kit (brand: Thermo, Cat. No: A15903). The PR receptor agonist Progesterone (brand: Sigma, Cat. No: P0130) was diluted 10-fold with DMSO in a 96-well V-bottom plate. The highest concentration was 100 μM, 8 concentrations in total. The test compound was diluted 10-fold with DMSO. The highest concentration was 3000 μM, 8 concentrations in total. The compound was then further diluted 50-fold with the detection buffer Nuclear Receptor Buffer F (containing 5 mM DTT) provided in the kit, 10 μL of the diluted compound was transferred to a 96-well half-area microplate, 5 μL of PR-LBD protein (4× concentration) was added to the test compound, then a mixture of 5 μL of fluorescein-coactivator peptide and Tb-labeled anti-GST antibody (4× concentration) was added, and the experiment was performed in duplicate. The plate was incubated at room temperature for 2 h, the fluorescence values (Excitation 337, Emission 520/495 nm) were detected using the PHERAstar instrument, and the 520:495 ratio was calculated. With the log value of the final concentration of the compound as the X axis and the 520/495 ratio as the Y axis, data was input into the processing software Graphpad Prism 9, and the EC50 value was calculated by four-parameter fitting. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |