PDE11A4 Assay Kit

The PDE11A4 Assay Kit is a fluorescence polarization (FP), homogeneous, 96-well assay kit designed for the screening and profiling of PDE11A4 (Phosphodiesterase 11A4) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant human PDE11A4, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 100 reactions.Figure 1: Illustration of the PDE11A4 Assay Kit principle.The assay uses a fluorescein-labeled cyclic adenosine monophosphate (cAMP-FAM for PDE11A4), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP) . PDE11A4 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 PDE11A4 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 PDE11A4? Check out ourPhosphodiesterase Screening Services.

Product Specifications

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 PDE4,7 and 8 being cAMP-specific hydrolases, and thus regulating positive and negative responses to it. PDE11A4 refers to a specific isoform of the phosphodiesterase 11A (PDE11A) enzyme. This enzyme is part of a family that break down cAMP and cGMP. The role of PDE11A, including its isoform PDE11A4, is crucial in regulating cellular signaling by controlling the levels of these cyclic nucleotides, which are involved in various physiological processes. These molecules play an important role in controlling various cellular functions, including cell growth, differentiation, and metabolism. By controlling the degradation of cAMP and cGMP, PDE11A4 influences several key processes like the activity of protein kinases, ion channels, and gene expression. PDE11A is highly expressed in the brain, particularly in areas like the cortex, hippocampus, and basal ganglia. PDE11A4, as an isoform, plays a role in regulating neurotransmitter systems that involve cAMP and cGMP. It is involved in processes like memory, learning, and mood regulation, and dysfunction in PDE11A or its isoforms could contribute to neurodegenerative disorders, cognitive impairments, or even mental health conditions.

UniProt

Q9HCR9

Applications

Study enzyme kinetics and screen small molecule inhibitors for drug discovery in high throughput screening (HTS) applications.

Format

Catalog #NameAmountStorage60110PDE11A4, GST-Tag*˃1 µg-80°C60200FAM-Cyclic-3′, 5′-AMP**1.2 nmoles**-80°C60393PDE Assay Buffer (Incomplete) 25 ml-20°C60390PDE Binding Agent100 µl4°C60391Binding Agent Diluent (cAMP) 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′-AMP is provided as a powder. Vial will need to be resuspended in 600 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.It is recommended that the compound alone is tested to determine any potential interference of the compound on the assay results.