| Assay Method Information | |
| | IMAP-FP Assay |
| Description: | The inhibitory activity of the representative compounds according to the present invention against phosphodiesterase 9A was tested based on the polarization assay (IMAP-FP screening express kit) provided by MDS (MDS Analytical Technologies, Sunyvale, CA, USA). As a buffer solution, a reaction solution (a reaction solution containing 0.01% Tween 20 provided by MDS was diluted 5 times, then 5 mM DTT is added) and a polarization detection solution (binding solution A and B provided by MDS were mixed at a ratio of 3:1 and IMAP binding reagent 1/600 was added) were prepared. 100 μM of the substrate (Fl-cGMP substrate; MDS) and 3.6 μg/ml of the enzyme PDE9A (ab54113; abeam) were prepared. 3.6 μg/ml PDE9A and 100 μM substrate were diluted to 20 ng/ml (final reaction concentration: 5 ng/ml) and 400 nM (final reaction concentration: 100 nM), respectively. The buffer solution used for all dilution and preparation processes was 1× reaction solution with 1 mM DTT added, and the polarization detection solution was used to induce polarization at the end. |
| Affinity data for this assay | |
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