PDE6C Assay Kit

Products from BPS Bioscience require a minimum order value above 400€

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

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. PDE6C, also known as cGMP-inhibited phosphodiesterase, is involved in light detection and cone phototransduction. PDE6C encodes the alpha-prime subunit of cone-specific phosphodiesterase, an enzyme found exclusively in the light-detecting (photoreceptor) cells called cones. Cone cells are found in a specialized tissue at the back of the eye known as the retina, and mutations in PDE6c are associated with eye diseases including Cone Dystrophy 4 and Achromatopsia. Continuing to understand PDE6C roles and the development of newer specific therapeutic tools will open new treatment avenues in PDE-related disorders.

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.

Description: The PDE6C Assay Kit is a fluorescence polarization (FP), homogeneous, 96-well assay kit designed for the screening and profiling of PDE6C (Phosphodiesterase 6C) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant PDE6C, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 100 reactions.The assay uses a fluorescein-labeled cyclic guanosine monophosphate (cGMP-FAM for PDE6C), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP). PDE6C 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 PDE6C 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 July 2024, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.

Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed

Supplied As: The PDE6C inhibitor screening assay kit comes in a convenient 96-well format, with purified PDE6C enzyme, fluorescently labeled PDE6 substrate (cGMP), binding agent, and PDE assay buffer for 100 enzyme reactions

Uniprot: P51160

Warnings: Avoid freeze/thaw cycles. 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.

Biosafety Level: Not applicable (BSL-1)

References: Ahmed W., et al., 2021 Biomedicine & Pharmacotherapy 134:111128.
Huang W., et al., 2020 Nat Commun 11:3191.

• Artikelnummer: BPS79501-1
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 79501-1
• Verpackungseinheit: 96 reactions
• Mengeneinheit: PAK