| Assay Method Information | |
| | Inhibitory Activity of Compound 1 on Phosphodiesterase 4B Subtype (PDE4B Enzyme) |
| Description: | Procedures:1. Dissolving the recombinant human PDE4B enzyme and enzyme substrate (1 μM cAMP) in freshly prepared assay buffer, respectively;2. Transferring the above PDE4B enzyme buffer solution to reaction wells;3. Adding compound 1 dissolved with 100% DMSO to reaction wells with the PDE4B enzyme buffer solution via acoustic technology (echo 550 nanoliter range) and incubating at room temperature for 10 min;4. Then, adding the enzyme substrate buffer solution to the above reaction wells to initiate reaction;5. Incubating at room temperature for 1 h;6. Adding the detection mixture (Transcreener® AMP2/GMP2 antibody and AMP2/GMP2 AlexaFluor633 tracing) to terminate the reaction and incubating with slow mixing for 90 min. The range of fluorescence polarization determination was Ex/Em=620/688.Data Analysis: The fluorescence polarization signal was converted into nM based on AMP/GMP standard curve and % enzyme activity calculated by Excel software relative to the DMSO control. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |