| Assay Method Information | |
| | Radiolabel Binding Studies for the Sigma-2 Receptor |
| Description: | The canine PDE3A assay kit was purchased from BPS Bioscience (San Diego, CA) which included dog PDE3A recombinant enzyme, FAM-cyclic-3',5'-AMP (20 uM), PDE3 assay buffer, binding agent, binding agent diluent and microtiter 96-well plates. Compounds were initially dissolved in DMSO. The stock concentration was subsequently diluted in cPDE assay buffer (10 stock) and serially diluted to give a concentration response curve starting at 10 uM (>7 total concentrations). Following the serial dilution, compound solution (0.1% final DMSO) was transferred to an assay plate (96-well) where the remaining canine PDE3A assay reagents were dispensed according to the BPS Bioscience protocol (catalog #79735) procedure using a final concentration of 5 pg/uL of enzyme. Each compound was evaluated for the inhibition of the canine PDE3A enzyme activity using the measurement of fluorescent polarization changes. The 50% inhibition concentration (IC50) nM was calculated and reported. As a comparison to the current cPDE3 inhibitors of the invention, pimobendan had a cPDE3 IC50 value of about 1530 nM. |
| Affinity data for this assay | |
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