| Assay Method Information | |
| | Inhibitory Effect of Compounds on BRD |
| Description: | Homogeneous time=resolved fluorescence (HTRF) was used to detect the binding of the compound to BRD4(D1 + D2) and BRDT (D1) proteins, and the AlphaScreen method used to detect the binding of the compound to BRD2 (D1+D2) and BRD3 (D1+D2) proteins. Detection steps: 1) All compounds were gradiently diluted on Echo plate according to the arrangement of the test plate. The final concentration of DMSO was 0.1%. 2) Compounds or DMSO was transferred to a 384-well assay plate by using Echo autosampler. 3) 2x concentration of protein and peptide mixture was added to the assay plate. 4) 2x concentration of the mixed detection solution was added to the assy plate and shaken for 30 seconds. 5) The plate was incubated at room temperature for 2 hours. 6) The fluorescence signal was read on Envision microplate reader (with excitation light wavelength at 349nM and emission light wavelengths at at 615 nm and 665 nm). 7) The curve was fitted. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |