PDELight HTS cAMP Phosphodiesterase Assay Kit
- Simple, Homogeneous Assay - add reagents and read
- Non-radioactive - no toxic reagents
- Rapid: 384-well plate in 15 min
- Bioluminescent sensitivity
- Low false positive rates
Phosphodiesterases (PDEs) function in conjunction with adenylate cyclase to regulate the amplitude of the ubiquitous second messenger signalling molecule, cyclic adenosine monophosphate (cAMP). Phosphodiesterases hydrolyse cAMP to adenosine monophosphate (AMP).
The PDELight™ assay is a non-radioactive, bioluminescent detection system for the activity of phosphodiesterases which utilise cAMP as their preferential second messenger. The AMP produced from the hydrolysis of cAMP is quantified using the PDELight™ AMP detection reagent which converts this AMP directly to ATP. The assay uses luciferase, which catalyses the formation of light from the newly formed ATP and luciferin. The emitted light is directly proportional to the level of AMP present in the reaction.
The assay can be performed with any cAMP dependant phosphodiesterase and unlike other assays does not require radioactive substrates, beads, modified substrates or antibodies. The assay can be performed in 96 or 384 well formats and is suitable for miniaturization to 1536 well plates or beyond. The assay can be easily and rapidly optimized for each phosphodiesterase, with linearity over 20µM AMP, a half life in excess of 2 hours and excellent Z’ values. It easily detects known phosphodiesterase inhibitors and provides an easily determined end point for IC50 analysis.