| Assay Method Information | |
| | In Vitro Detection of the Inhibitory Activity of Compounds Against PDE 4B Enzyme |
| Description: | Procedure:1. The recombinant human PDE4B enzyme and enzyme substrate (1 uM cAMP) were each dissolved in newly prepared experimental buffer solution;2. The PDE4B enzyme buffer solution was transferred into reaction wells;3. The compound which was dissolved in 100% DMSO was added to the reaction wells containing PDE4B enzyme buffer solution by acoustic technique (echo 550; millilambda range) and the mixture was incubated for 10 minutes at room temperature;4. The enzyme substrate buffer solution was added to the above reaction wells to initiate reaction;5. The resulting mixture was incubated at room temperature for 1 hour;6. The detection mixture (Transcreener AMP2/GMP2 antibody and AMP2/GMP2 AlexaFluor633 tracer) was added to stop the reaction, and the resulting mixture was incubated for 90 minutes while slowly mixing. The measurement range of fluorescence polarization was Ex/Em=620/688. |
| Affinity data for this assay | |
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