| Assay Method Information | |
| | Effect of Compounds on PDE4B2 Activity Assay |
| Description: | The effect of compounds on PDE4B2 activity was detected using a fluorescence polarization kit (BPS Bioscience, Catalog #60343). According to the instructions of the kit, FAM-Cyclic-3′,5′-AMP at a final concentration of 0.1 M, 1 ng/well of PDE4B2 (for a negative control, a PDE buffer was added), and gradient-diluted compounds (for a positive control well, a PDE buffer containing 10% of DMSO was added) were added to each well, fully mixed, and then reacted at room temperature for 1 h. The Binding Agent was diluted with the Binding Agent Diluent (cAMP) at a ratio of 1:100 for later use, the Binding Agent dilution was taken and added to the test plate at 50 μl/well and incubated at room temperature with slow shaking for 20 minutes. After the incubation was complete, Envision was used for FP detection, with Excitation at 480 nm and Emission at 535 nm. |
| Affinity data for this assay | |
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