| Assay Method Information | |
| | Fluorescence Polarization Assay |
| Description: | For KD determinations, 10 nM JQ1-TAMRA probe was incubated in 384-well smallvolume black microtiter plates with serial (1:2) dilutions of each BRD4 variant (0.61 nM to 10 μM) in a buffer containing 10 mM HEPES, pH 7.5 (Applichem), 150 mM NaCl (Sigma), 0.005% Tween 20 (Sigma), and 0.5 mM Tris(2-carboxyethyl)phosphine(Calbiochem). After 30 min of incubation at room temperature, the plates were measured in a Tecan M1000 plate reader using its FP module and excitation/emission wavelengths of 530 and 570 nm, respectively. |
| Affinity data for this assay | |
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