| Description: | The PDE Assay Kit is designed for identification of PDE inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDEs to the binding agent. The key to the PDE Assay Kit is the spe |
|---|---|
| Background: | Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. Phosphodiesterases catalyze the hydrolysis of the phosphodiester bond in dye-labeled cyclic monophosphates. Beads selectively bind the phosphate group in the nucleotide product. This increases the size of the nucleotide relative to unreacted cyclic monophosphate. In the polarization assay, dye molecules with absorption transition vectors parallel to the linearly-polarized excitation light are selectively excited. Dyes attached to the rapidly-rotating cyclic monophosphates will obtain random orientations and emit light with low polarization. Dyes attached to the slowly-rotating nucleotide-bead complexes will not have time to reorient and therefore will emit highly polarized light. |
| Synonym(s): | PDE, phosphodiesterase, universal |
| Supplied As: | The PDE inhibitor screening assay kit comes in a convenient 96-well format, with fluorescently labeled substrate (cAMP and cGMP), binding agent, and PDE assay buffer for 100 enzyme reactions. |
| Contraindications: | DMSO >1%, strong acids or bases, ionic detergents, high salt |
| Format: | |
| Instructions for use: | See assay kit data sheet for detailed protocol. |
| Storage / Stability: | At least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival. |
| Application(s): | Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications. |
| Reference(s): | Maurice DH. Front Biosci. 2005; 10:1221-8. |
| Notes: | MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED: Fluorescent microplate reader capable to measure fluorescence polarization |
| Warning(s): | Avoid freeze/thaw cycles |
| Scientific Category: | Phosphodiesterase(s) |
| Product Type: | Assay Kits |