| Assay Method Information | |
| | Inhibitory Effect of the Compound According to the Present Invention on BRD |
| Description: | The compounds were screened in vitro, and each compound was serially diluted to 10 concentrations. Four proteins BRD4 (D1+D2), BRDT (D1), BRD2 (D1+D2) and BRD3 (D1+D2) were chosen to determine the IC50 values of compounds (see Table 1).3. Experimental Materials:BRD2(1,2)(BPS, Cat. No. 31024)BRD3(1,2)(BPS, Cat. No. 31035)BRDT(D1)(Active Motif, Cat. No. 31450)BRD4(1,2)(BPS, Cat. No. 31044)(+)-JQ1(BPS, Cat. No. 27402)4. Compound treatment: The test compound was dissolved in dimethyl sulfoxide (DMSO), and the storage concentration was 10 mM.5. Steps for Homogeneous Time-Resolved Fluorescence Detection:1) According to the arrangement of the detection plate, all compounds were diluted in Echo plate.The final dilution concentration of DMSO is 0.1%.2) The compound or DMSO was transferred to a 384-well detection plate with the Echo automatic sampler.3) Two-fold concentration of the mixture of protein and peptide was added to the detection plate.4) Two-fold concentration of mixed detection solution was added to the test plate, and shaken for 30 s.5) Incubating for 2 h at room temperature.6) Fluorescence signal was read on Envision multi-function microplate reader (excitation light wavelength being 340 nm, and emission light wavelength being 615 nm and 665 nm). |
| Affinity data for this assay | |
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