| Assay Method Information | |
| | Pharmacodynamic Tests |
| Description: | In the present disclosure, a fluorescent molecular probe was constructed based on a MLL1 peptide fragment binding to WDR5, for use as a method for study on an aniline compound interfering with WDR5 protein-protein interaction, to determine an inhibition rate of the aniline compound at various concentrations, and then compute the IC50 value. Specific experimental steps: 20 μL of WDR5 protein, 20 μL of a fluorescent probe, and 20 μL of a compound of different concentration gradients were added to a 384-well plate respectively. After incubation for 0.5 hrs, the fluorescence was read on a multifunctional microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 535 nm, to compute the mP value. The inhibition rate was computed as per the formula below, and then the IC50 value was computed using GraphPad software. |
| Affinity data for this assay | |
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