Overview |
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Product |
PDEase KitTM |
Description |
PDEase KitTM is designed to allow investigators to determine PDE4 activity |
Contents |
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Incubation Buffer Reagent A |
5 mL |
Incubation Buffer Reagent B |
5 mL |
Incubation Buffer Reagent C |
5 mL |
PDE Substrate Buffer D |
10 mL |
PDE Substrate |
2X 200 µl |
Inhibitor Buffer |
50 mL |
Second Enzyme Source (10X) |
2X 250 µl |
Resin Slurry (10X) |
100 mL |
Control PDE4 Enzyme |
1x200 µl |
Introduction |
cAMP is a second messenger that is ubiquitously found in all mammalian cells. This messenger modulates the signal transduction of a variety of hormones, neurotransmitters, growth factors, cytokines and others ligand receptors. cAMP phosphodiesterase enzymes provide the sole pathway for hydrolyzing and truncating the cAMP signaling that is originated in response to receptor activation. The cAMP hydrolyzing activity is provided by a fairly large multi-enzyme family with distinct intracellular localization and regulatory properties. Among various PDE families (PDE1-PDE11), PDE4 has 4 genes (PDE4A, PDE4B, PDE4C, PDE4D) that are expressed in multi-splice variant forms.
The PDE4 activity measurement required a 2 step reaction, in the first reaction, cAMP is hydrolyzed by PDE activity in to non-cyclic form. This reaction is stopped by denaturing the PDE source or by addition of PDE4-selective inhibitor, followed by conversion of AMP to adenosine by the second enzymatic reaction to equilibrium. Since Adenosine is neutral, cAMP or AMP is separated by charge by ion exchange chromatography. In order to avoid background contribution of other PDEs, this PDE assay is carried out at substrate concentration optimized for PDE4. The PDE4 enzymatic activity is directly proportional to the adenosine formed in the first reaction.
This PDE4 activity measurement provides reproducible results within 2-3 hours. The kit provides enough reagents to process 50 or 100 samples in tandem along with internal controls. The kit reagents are stable for 6 months when stored at recommended conditions. The contents of this kit must be stored at different temperatures as indicated on the bottle labels and specification sheet.
Fabgennix PDEase Kit has been used in 9 Publications |
Joshi, Rashika, et al. "Phosphodiesterase (PDE) inhibitor torbafylline (HWA 448) attenuates burn-induced rat skeletal muscle proteolysis through the PDE4/cAMP/EPAC/PI3K/Akt pathway." Molecular and cellular endocrinology 393.1 (2014): 152-163. PMID24973766
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Al-Tawashi, Azza, and Chris Gehring. “Phosphodiesterase Activity Is Regulated by CC2D1A That Is Implicated in Non-Syndromic Intellectual Disability.” Cell Communication and Signaling?: CCS 11 (2013): 47. PMC3704924
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Gobejishvili, Leila et al. “Rolipram Attenuates Bile Duct Ligation–Induced Liver Injury in Rats: A Potential Pathogenic Role of PDE4.” The Journal of Pharmacology and Experimental Therapeutics 347.1 (2013): 80–90. PMC. PMC3781411
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Gobejishvili, Leila et al. “S-Adenosylmethionine Decreases Lipopolysaccharide-Induced Phosphodiesterase 4B2 and Attenuates Tumor Necrosis Factor Expression via cAMP/Protein Kinase A Pathway.” The Journal of Pharmacology and Experimental Therapeutics 337.2 (2011): 433–443. PMC3083110
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Li, Yuting et al. “Regulation of Amygdalar PKA by β-Arrestin-2/phosphodiesterase-4 Complex Is Critical for Fear Conditioning.”Proceedings of the National Academy of Sciences of the United States of America 106.51 (2009): 21918–21923. PMC2799795
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Gobejishvili, Leila et al. “Enhanced PDE4B Expression Augments LPS-Inducible TNF Expression in Ethanol-Primed Monocytes: Relevance to Alcoholic Liver Disease.” American Journal of Physiology - Gastrointestinal and Liver Physiology 295.4 (2008): G718–G724. PMC2575909
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Kim, Jong-So, et al. "Daily rhythm in pineal phosphodiesterase (PDE) activity reflects adrenergic/3′, 5′-cyclic adenosine 5′-monophosphate induction of the PDE4B2 variant." Endocrinology 148.4 (2007): 1475-1485. PMID17204557
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Janciauskiene, Sabina M., Izabela M. Nita, and Tim Stevens. "α1-Antitrypsin, Old Dog, New Tricks α1-ANTITRYPSIN EXERTS IN VITRO ANTI-INFLAMMATORY ACTIVITY IN HUMAN MONOCYTES BY ELEVATING cAMP." Journal of Biological Chemistry 282.12 (2007): 8573-8582. PMID17261591
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Richard, François, et al. "Methods, Compositions and Kits for the Evaluation and Preservation of Sperm." U.S. Patent Application 13/365,838. 13/365,838
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Publishing research using PDEase Kit? Please let us know, we can cite the reference in this datasheet.
Product | Note | Status | Price | |
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FGI-1943 | |||
Display accessory details |
PD4-101AP
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PD4A-112AP
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PD4C-301AP
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PD4-401AP
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