| Assay Method Information | |
| | Inhibition Assay |
| Description: | The PDE10 inhibition assay in 384-well plates was conducted to identify substances for the inhibition of cyclic nucleotide hydrolysis by the PDE10 enzyme. The cyclic nucleotide substrate concentration used was at a Km concentration (25 nM final). PDE10 activity was measured using Scintillation Proximity Assay (SPA)-based methods. PDE10 catalyses the hydrolysis of the intracellular messenger adenosine 5'-cyclic phosphate (cAMP) to the non-cyclic adenosine 5'-monophosphate (AMP). The SPA assay was based upon the selective interaction of the tritiated product with yttrium oxide LEADseeker beads.The assay was performed in 10 uL samples containing 5 uL of 0.3 ng/mL PDE10 final and 5 uL of 3H-5' cAMP (PerkinElmer, NET111540) run at Km of 25 nM final. The assay buffer contained 25 mM HEPES pH 7.4, 2.5 mM Magnesium Chloride and 0.1% BSA. Compound dose response curves were pre-incubated with 5 L of 2PDE10 enzyme for 10 minutes. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |