Products from BPS Bioscience require a minimum order value above 400€Application: Useful for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and 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. PDE4 is the second most abundant PDE in cardiovascular tissues. PDE4, is a cAMP-specific PDE and the largest PDE subfamily with over 35 different isoforms and as such, the most widely characterized PDE isoenzyme. It is the predominant isoenzyme in most inflammatory cells, and the airways smooth muscle and is implicated in inflammatory airways disease. PDE4 inhibitor roflumilast has been approved for the treatment of chronic obstructive lung disease (COPD).
Description: The PDE4C1 Assay Kit is a fluorescence polarization (FP), homogeneous, 96-well assay kit designed for the screening and profiling of PDE4C1 (Phosphodiesterase 4C1) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant PDE4C1, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 100 or 400 reactions. The assay uses a fluorescein-labeled cyclic guanosine monophosphate (cAMP-FAM for PDE4C1), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP). PDE4C1 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 PDE4C1 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 January, 2025, 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 PDE4C1 inhibitor screening assay kit comes in a convenient 96-well format, including purified PDE4C1 enzyme, fluorescently labeled PDE4C1 substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
Uniprot: Q08493
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 (bpsbioscience.com/fluorescence-polarization-assays-principles-applications ) . Avoid freeze/thaw cycles.
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.
English
Deutsch
