DESCRIPTION

• Shiptemp: -80°C (dry ice)
• Synonyms: inhibitor screening, assay kit, PDE7B mouse, pde7b
• Warnings: Avoid freeze/thaw cycles.
• Category: Phosphodiesterase/Assay Kit
• Background: Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE7B is highly expressed in the pancreas and in other various tissues. Inhibition of PDE7B leads to induction of apoptosis. 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.
• References: 1. Brooks, M.D., Jackson, E., et al. PDE7B Is a Novel, Prognostically Significant Mediator of Glioblastoma Growth Whose Expression Is Regulated by Endothelial Cells. PLOS ONE. 2014 9(9): e107397. 2. Gardner, C., Robas, N., et al. Cloning and Characterization of the Human and Mouse PDE7B, a Novel cAMP-Specific Cyclic Nucleotide Phosphodiesterase, Biochemical and Biophysical Research Communications, 2000, 272(1): 186-192.
• Description: The Mouse PDE7B Assay Kit is designed for identification of inhibitors of Mouse PDE7B using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by Mouse PDE7B to the binding agent. The key to the Mouse PDE7B Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for Mouse PDE7B reactions. First, the fluorescently labeled cAMP is incubated with a sample containing Mouse PDE7B for 1 hour. Second, a binding agent is added to the reaction mix to produce a change in fluorescent polarization that can then be measured using a fluorescence reader equipped for the measurement of fluorescence polarization.
• Methodology: Fluorescence Polarization
• Supplied As: The Mouse PDE7B Assay Kit comes in a convenient 96-well format, with purified Mouse PDE7B enzyme, fluorescently labeled substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
• Unspsc Code: 41121811
• Unspsc Name: Laboratory microchemistry kits
• Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
• Product Type: Assay Kit
• Biosafety Level: Not applicable (BSL-1)
• Related Products: 60371, 60075, 60073, 60412, 60690-1, 60182, 60185, 79534, 60690
• Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
• 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.
• Scientific Category: Phosphodiesterase
• Instructions for Use: See assay kit datasheet for detailed protocol.