PDE11A4 Assay Kit
The PDE11A4 Assay Kit is designed for identification of PDE11A4 inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE11A4 to the binding agent. The key to the PDE11A4 Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE11A4 reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE11A4 for 1 hour. Second, a binding agent is added to the reaction mix to produce a change in fluorescent polarization. The FP signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.
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Catalog # |
Component |
Amount |
Storage |
|
>1 µg |
-80°C |
(Avoid freeze/ thaw cycles!) |
||
50 µl |
-80°C |
|||
-20°C |
||||
100 µl |
+4°C |
|||
10 ml |
+4°C |
|||
79685 |
Black, low binding, microtiter plate |
1 |
Room temp. |
Phosphodiesterases (PDEs) are involved in the dynamic regulation of cAMP and cGMP signaling. PDE11A4 has been observed in skeletal muscle, prostate, testis, salivary gland, thyroid gland, and liver. PDE11A4 may play role in the CNS with an association to major depressive disorder. 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.