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Assay Method Information

Assay Name:  BRD4-BD1 Binding Assay
Description:  11 compounds were evaluated against BRD4-BD1 in vitro with compound 28a as the reference compound. BRD4 (44-168 aa) was bought from Active Motif. After preparing 1 assay buffer (modified HEPES Buffer), target compounds were transferred to assay plate by Echo, and the final concentration of DMSO solution was 0.1%. Then the preparation of protein solution and substrate solution in 1 assay buffer was performed. After that, 5 L of protein solution was transferred to assay plate and incubated at room temperature for 15 min. The protein concentration was 5 nM. Then 5 L of substrate solution was added to each well to start reaction with an incubation period for 60 min at room temperature. Finally, 15 L acceptor and donor solution were added, followed by another incubation period for 60 min at room temperature under subdued light.
Affinity data for this assay
 

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Last update November 1, 2007
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