TAMRA-cAMP PDE IV substrate *Red Fluorescence*
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Additional ordering information
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Bulk request | Inquire |
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Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | ~900 |
Solvent | DMSO |
Spectral properties
Correction Factor (260 nm) | 0.32 |
Correction Factor (280 nm) | 0.178 |
Extinction coefficient (cm -1 M -1) | 90000 |
Excitation (nm) | 552 |
Emission (nm) | 578 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Overview | SDSProtocol |
See also: Cyclic AMP (cAMP) Signaling
Molecular weight ~900 | Correction Factor (260 nm) 0.32 | Correction Factor (280 nm) 0.178 | Extinction coefficient (cm -1 M -1) 90000 | Excitation (nm) 552 | Emission (nm) 578 |
This red cAMP derivative is a specific substrate for phosphodiesterase (PDE) IV. It can be used for assaying PDE IV activities or screening PDE IV inhibitors in combination with anti-cAMP antibody in a FRET readout or FP format. PDE is a group of enzymes that degrade the second messenger molecules: cyclic nucleotides cAMP and cGMP. They regulate the localization, duration, and amplitude of cyclic nucleotide signaling within subcellular domains. PDEs are therefore important regulators of signal transduction mediated by these second messenger molecules. PDE enzymes are often targets for pharmacological inhibition due to their unique tissue distribution, structural and functional properties. Inhibitors of PDE can prolong or enhance the effects of physiological processes mediated by cAMP or cGMP by inhibition of their degradation by PDE. PDE inhibitors have been identified as new potential therapeutics in areas such as pulmonary arterial hypertension, coronary heart disease, dementia, depression and schizophrenia.
Platform
Fluorescence microplate reader
Excitation | 540 nm |
Emission | 590 nm |
Cutoff | 570 nm |
Recommended plate | Solid black |
Example protocol
AT A GLANCE
Important notes
Following protocol only provides a guideline, and should be modified according to your specific needs.
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1. TAMRA-cAMP PDE IV stock solution (1 mM):
Make a 1 mM stock solution by adding 500 µL of DMSO into the vial of 0.5 umol TAMRA-cAMP PDE IV substrate. Note: Unused stock solution can be stored at -20 oC in dark place in single aliquotes.
PREPARATION OF WORKING SOLUTION
TAMRA-Cyclic-3’, 5’-AMP PDE IV substrate assay solution (2X):
Make 2X TAMRA-Cyclic-3’, 5’-AMP PDE IV substrate assay solution by diluting 1 mM TAMRA-Cyclic-3’ ,5’-AMP PDE IV substrate stock solution into your PDE buffer (such as 10 mM Tris-HCl, pH 7.4, 10 mM Mg Cl2, 1 mM MnCl2) to make a 200 - 400 nM solution. Note: Make only sufficient quantity needed for the assay.
SAMPLE EXPERIMENTAL PROTOCOL
- Mix equal volume of the PDE IV standards or samples with 2X TAMRA-Cyclic-3’ ,5’-AMP PDE IV substrate assay solution, and incubate at room temperature for at least 1 hour.
- Monitor the fluorescence polarization at Ex/Em = 540/590 nm.
Spectrum
Open in Advanced Spectrum Viewer
Spectral properties
Correction Factor (260 nm) | 0.32 |
Correction Factor (280 nm) | 0.178 |
Extinction coefficient (cm -1 M -1) | 90000 |
Excitation (nm) | 552 |
Emission (nm) | 578 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Correction Factor (260 nm) | Correction Factor (280 nm) |
FAM-cAMP PDE IV substrate *Green Fluorescence* | 493 | 517 | 83000 | 0.32 | 0.178 |
References
View all 34 references: Citation Explorer
Oxidative stress employs phosphatidyl inositol 3-kinase and ERK signalling pathways to activate cAMP phosphodiesterase-4D3 (PDE4D3) through multi-site phosphorylation at Ser239 and Ser579
Authors: Hill EV, Sheppard CL, Cheung YF, Gall I, Krause E, Houslay MD.
Journal: Cell Signal (2006): 2056
Authors: Hill EV, Sheppard CL, Cheung YF, Gall I, Krause E, Houslay MD.
Journal: Cell Signal (2006): 2056
Leptin interferes with adrenocorticotropin/3',5'-cyclic adenosine monophosphate (cAMP) signaling, possibly through a Janus kinase 2-phosphatidylinositol 3-kinase/Akt-phosphodiesterase 3-cAMP pathway, to down-regulate cholesterol side-chain cleavage cytochr
Authors: Hsu HT, Chang YC, Chiu YN, Liu CL, Chang KJ, Guo IC.
Journal: J Clin Endocrinol Metab (2006): 2761
Authors: Hsu HT, Chang YC, Chiu YN, Liu CL, Chang KJ, Guo IC.
Journal: J Clin Endocrinol Metab (2006): 2761
Helix-1 of the cAMP-specific phosphodiesterase PDE4A1 regulates its phospholipase-D-dependent redistribution in response to release of Ca2+
Authors: Huston E, Gall I, Houslay TM, Houslay MD.
Journal: J Cell Sci (2006): 3799
Authors: Huston E, Gall I, Houslay TM, Houslay MD.
Journal: J Cell Sci (2006): 3799
TcPDE4, a novel membrane-associated cAMP-specific phosphodiesterase from Trypanosoma cruzi
Authors: Alonso GD, Schoijet AC, Torres HN, Flawia MM.
Journal: Mol Biochem Parasitol (2006): 40
Authors: Alonso GD, Schoijet AC, Torres HN, Flawia MM.
Journal: Mol Biochem Parasitol (2006): 40
Intracellular targeting of phosphodiesterase-4 underpins compartmentalized cAMP signaling
Authors: Lynch MJ, Hill EV, Houslay MD.
Journal: Curr Top Dev Biol (2006): 225
Authors: Lynch MJ, Hill EV, Houslay MD.
Journal: Curr Top Dev Biol (2006): 225
Compartmentalization of cAMP-Dependent Signaling by Phosphodiesterase-4D Is Involved in the Regulation of Vasopressin-Mediated Water Reabsorption in Renal Principal Cells
Authors: Stefan E, Wiesner B, Baillie GS, Mollajew R, Henn V, Lorenz D, Furkert J, Santamaria K, Nedvetsky P, Hundsrucker C, Beyermann M, Krause E, Pohl P, Gall I, Macintyre AN, Bachmann S, Houslay MD, Rosenthal W, Klussmann E.
Journal: J Am Soc Nephrol. (2006)
Authors: Stefan E, Wiesner B, Baillie GS, Mollajew R, Henn V, Lorenz D, Furkert J, Santamaria K, Nedvetsky P, Hundsrucker C, Beyermann M, Krause E, Pohl P, Gall I, Macintyre AN, Bachmann S, Houslay MD, Rosenthal W, Klussmann E.
Journal: J Am Soc Nephrol. (2006)
PDE7A1, a cAMP-specific phosphodiesterase, inhibits cAMP-dependent protein kinase by a direct interaction with C
Authors: Han P, Sonati P, Rubin C, Michaeli T.
Journal: J Biol Chem (2006): 15050
Authors: Han P, Sonati P, Rubin C, Michaeli T.
Journal: J Biol Chem (2006): 15050
Importance of cAMP-response element-binding protein in regulation of expression of the murine cyclic nucleotide phosphodiesterase 3B (Pde3b) gene in differentiating 3T3-L1 preadipocytes
Authors: Liu H, Tang JR, Choi YH, Napolitano M, Hockman S, Taira M, Degerman E, Manganiello VC.
Journal: J Biol Chem (2006): 21096
Authors: Liu H, Tang JR, Choi YH, Napolitano M, Hockman S, Taira M, Degerman E, Manganiello VC.
Journal: J Biol Chem (2006): 21096
DdPDE4, a novel cAMP-specific phosphodiesterase at the surface of dictyostelium cells
Authors: Bader S, Kortholt A, Snippe H, Van Haastert PJ.
Journal: J Biol Chem (2006): 20018
Authors: Bader S, Kortholt A, Snippe H, Van Haastert PJ.
Journal: J Biol Chem (2006): 20018
Phosphodiesterase-5 Gln817 is critical for cGMP, vardenafil, or sildenafil affinity: its orientation impacts cGMP but not cAMP affinity
Authors: Zoraghi R, Corbin JD, Francis SH.
Journal: J Biol Chem (2006): 5553
Authors: Zoraghi R, Corbin JD, Francis SH.
Journal: J Biol Chem (2006): 5553
Application notes
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A Non-Radioactive Photometric Assay for Glucose Uptake in Insulin-Responsive 3T3-L1 Adipocytes
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A Non-Radioactive Photometric Assay for Glucose Uptake in Insulin-Responsive 3T3-L1 Adipocytes
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Evaluation of FLIPR Calcium Assays for Screening GPCR and Calcium Channel Targets
Restriction of Advanced Glycation End Products Improves Insulin Resistance in Human Type 2 Diabetes
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Do you have a ready-to-use cell viability assay kit?
Is there a multi-drug resistance assay kit that can detect both P-glycoprotein (P-gp, MDR1) and the multidrug resistance associated protein (MRP1)?
What assay kits measure NAD/NADH from cell samples?