PDE9A2 Assay Kit

• Background :

  Phosphodiesterases (PDEs) play an important role in the dynamic regulation of the second messengers cAMP (cyclic adenosine monophosphate) and cGMP (cyclic guanosine monophosphate) signaling, by hydrolyzing them. The PDE superfamily is composed of 11 families, with PDE5 and PDE9 being cGMP-specific hydrolases, and thus regulating positive and negative responses to it. PDE9A2 is expressed in certain regions of the brain, where it plays a role in regulating synaptic transmission, learning, and memory. Its influence on cGMP levels can affect neuronal signaling. Since cGMP is a mediator in blood vessel relaxation, PDE9A2 has an impact on blood pressure regulation and overall vascular function.. Dysregulation of PDE9A2 may contribute to various conditions, including cardiovascular diseases, neurological disorders, and possibly cancer, as its activity influences cell signaling and growth. PDE9A2 is being explored as a target for drug development, particularly for neurological disorders like Alzheimer's disease (AD), as modulating its activity can impact cognitive functions. Inhibiting PDE9A2 could increase cGMP levels, which may have beneficial effects in neurodegenerative diseases.

• Description :

  The PDE9A2 Assay Kit is a fluorescence polarization (FP), homogeneous, 96-well assay kit designed for the screening and profiling of PDE9A2 (Phosphodiesterase 9A2) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant human PDE9A2, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 100 reactions.Figure 1: Illustration of the PDE9A2 Assay Kit principle.The assay uses a fluorescein-labeled cyclic guanosine monophosphate (cGMP-FAM for PDE9A2), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP) . PDE9A2 catalyzes the hydrolysis of the phosphodiester bond in the cyclic nucleotide and frees the phosphate group. In a second step the free phosphate group is recognized by a specific phosphate-binding nanobead (Binding Agent) leading to the formation of a large complex, with restricted movement (high FP) . FP is proportional to PDE9A2 activity.This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note:FP, assay principles and applications.Note:As of February 2025, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.Need us to run inhibitor screens or profile your compounds against PDE9A2? Check out ourPhosphodiesterase Screening Services.

• UniProt :

  O76083

• Applications :

  Useful for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.

• Format :

  Catalog #NameAmountStorage60090PDE9A2, GST-Tag*˃1 µg-80°C60201FAM-Cyclic-3′, 5′-GMP**2 x 1.2 nmoles**-80°C60393PDE Assay Buffer (Incomplete) 25 ml-20°C60390PDE Binding Agent100 µl4°C60392Binding Agent Diluent (cGMP) 10 ml4°C827350.5 M DTT200 µl-20°C79685Low binding, black 96-well plate1Room Temp.* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.**FAM-Cyclic-3′, 5′-GMP is provided as a powder. Vial will need to be resuspended in 400 µl of Complete PDE Assay Buffer before use.

• Shipping Conditions :

  -80°C

• Storage Conditions :

  This assay kit will perform optimally for up to6 monthsfrom date of receipt when the materials are stored as directed.

• Contraindications :

  The final concentration of DMSO in the assay should not exceed 1%.Fluorescent compounds that have λex=470 nm and detection at λem=528 nm can interfere with the readouts.