| Assay Method Information | |
| | Inhibition Assay |
| Description: | Compounds were tested for their activity by measuring the inhibition of PDE7A or PDE7B hydrolysis of [3H]cAMP to [3H]AMP. Generally, eight dilutions of compound were assayed in 50 mM Tris-HCl pH 7.5, 8.3 mM MgCl2, 0.5 mg/mL BSA, 1.7 mM EGTA, 16 nM [3H]cAMP, and 1% DMSO. PDE7A or PDE7B (typically 1-5 ng/mL) (BPS BioSciences, CA) is added to initiate the reaction. The reaction was incubated at 30° C. for 20 minutes. PDE7A and PDE7B hydrolysis was terminated by the addition of yttrium silicate beads (GE Healthcare, RPNQ0150) and counted on a Wallac Microbeta scintillation counter, 1-2 hours following the addition of the beads. |
| Affinity data for this assay | |
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