PDEase Kit

Product no.: PDEase-100

$425.00

Description
Publications

Overview
Product	PDEase KitTM
Description	PDEase KitTM is designed to allow investigators to determine PDE4 activity
Contents
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.

PDEase Kit Protocol

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

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

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

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

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

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

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

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

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

Publishing research using PDEase Kit? Please let us know, we can cite the reference in this datasheet.

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