| Assay Method Information | |
| | Binding Inhibitory Test on BRD3 Protein |
| Description: | The binding inhibitory effect of the compounds of the present invention on the BRD3 protein among the BET proteins was tested as follows and confirmed. The compounds were diluted 1:5 serial dilutions in an assay buffer from a 10 mM stock in DMSO in a white OptiPlate. A mixture consisting of 100 nM GST BRD2 (BD1, BD2, BD1+BD2) and 100 nM biotinylated acetyl histone H4 (Lys5, 8, 12, 16) peptides was added to the diluent, and then each sample was incubated with shaking at 300 rpm for 30 minutes at room temperature in the dark. Thereafter, signals were measured with a PerkinElmer Envision HTS Multilabel Reader using a PerkinElmer Alpha Screen protocol. Determination of IC50 values was performed using GraphPad Prism 3.03 software. |
| Affinity data for this assay | |
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