| Assay Method Information | |
| | TR-FRET Assay |
| Description: | A TR-FRET assay was used to assess the effect of compounds disclosed herein on the interaction between RBM39 and the DCAF15 complex. Test compounds were dissolved to form a 10 mM DMSO stock solution. A 45 μL aliquot of the stock solution was transferred to a 384 pp-plate, and a threefold, 8-point dilution was performed via transferring 15 μL compound solution into 30 μL of DMSO. The plates were then spun at room temperature at 1,000 RPM for 1 minute. A 30 nL aliquot of the diluted compound was transferred to a 384 well plate, which was incubated at room temperature for 15 minutes. Next, Solutions 1, 2, and 3 were prepared as described in the below tables. A 5 μL aliquot of Solution 2 was added to each well, followed by 5 μL of Solution 3 to start the reaction. The final volume of each well was 10 μL. The plates were incubated at room temperature for 60 minutes, and then read. |
| Affinity data for this assay | |
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