| Assay Method Information | |
| | Cereblon Binding Experiment |
| Description: | The compounds of the present disclosure were dissolved in DMSO at a stock concentration of 10 mM. The dose-response program of the compound dilution and loading instrument (Echo) was used to perform gradient dilution of the compound stock solution. The total experimental volume of the dilution program was 20 μL, with an initial concentration of 100 μM for the compound to be tested and 200 μM for the standard. A 4-fold dilution was performed, resulting in 8 concentration points, and the DMSO content was 1%. After the program was completed, 5 μL of the 1×9 #diluent from the kit was added to each well, followed by the addition of 5 μL of the GST-labeled human Cereblon WT ligand. After thorough mixing, 10 μL of the HTRF detection reagent was added, and the mixture was incubated at room temperature for 3 h. The HTRF signal in each well was measured using an Envision plate reader. 100% binding inhibition was defined as the signal ratio under treatment with 200 μM standard lenalidomide. |
| Affinity data for this assay | |
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