Cyclic nucleotide phosphodiesterases (PDEs) are important drug targets with unique tissue distribution and functional properties. This family of enzymes is key in the regulation of cyclic nucleotide second messenger levels because they degrade both cAMP and cGMP. Some PDEs hydrolyze both cAMP and cGMP, while others are either cAMP or cGMP hydrolases. Selective PDE inhibitors block the activity of specific PDEs and are used to treat a variety of diseases. Non-selective PDE inhibitors include IBMX and caffeine, both of which are important consumables in basic research laboratories.
cAMP response to isoproterenol followed by inhibition of cAMP production with quinpirole. Detected with the cADDis cAMP assay.
Kinetic cGMP response to Sodium Nitroprusside. Detected with the GENIe cGMP assay.
PDE Selectivity Assay
An Optogenetic Tool for Monitoring PDE Rates
The blue-light activated adenylyl cyclase bPAC (#V0100N) can be co-expressed with R-cADDis (#U0200R), a red ﬂuorescent biosensor for cAMP. A two second pulse of blue light activates bPAC to stimulate cAMP production. cAMP degradation by PDE is monitored in real time using R-cADDis. Read about how this method can be used to study the role of PDE activity in neurodegeneration: Live-Cell Assays for Cell Stress Responses Reveal New Patterns of Cell Signaling Caused by Mutations in Rhodopsin, α-Synuclein and TDP-43.
Simple, step-by-step protocols for optimizing cAMP and cGMP assays are available online. These assays are non-FRET and cell lysis, enzymes, and co-factors are not necessary. Add our sensors to your cells, incubate, add drug, and measure fluorescence changes.
Sound Intriguing? Get more details on our PDE Biology Assays:
cADDis cAMP Assay
GENIe cGMP Assay
bPAC in BacMam
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