| Assay Method Information | |
| | Binding Activity of Compound of the Present Invention to Glucocorticoid Receptor GR |
| Description: | The binding activity of the compound to GR was determined using the Polarscreen Glucocorticoid Receptor Assay Kit, Red (brand: Thermo, Cat. No: A15898). The test compound was diluted 10-fold with DMSO in a 96-well V-bottom plate (brand: Nunc, Cat. No: 249944). The highest concentration was 100 μM, 8 concentrations in total. The compound was then further diluted 50-fold with the detection buffer Complete GR Screening buffer provided in the kit, 10 μL of the diluted compound was transferred to a 384 microplate (brand: Corning, Cat. No: 4514), 5 μL of Fluormone GS Red (4× concentration) was added to the test compound, then a mixture of 5 μL of GR Full length (4× concentration) was added, and the experiment was performed in duplicate. The 384 microplate were incubated at room temperature in the dark for 2 h, and the fluorescence polarization (mP) was detected using the En Vision multifunctional microplate reader (manufacturer: Perkinelmer). |
| Affinity data for this assay | |
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