logo
AAT Bioquest

TAMRA-cGMP PDE V substrate *Red Fluorescence*

Product Image
Product Image
Gallery Image 1
Ordering information
Price
Catalog Number
Unit Size
Quantity
Add to cart
Additional ordering information
Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
ShippingStandard overnight for United States, inquire for international
Request quotation
Physical properties
Molecular weight~1000
SolventDMSO
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-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


Molecular weight
~1000
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 cGMP derivative is a specific substrate for phosphodiesterase (PDE) V. It can be used for assaying PDE V activities or screening PDE V inhibitors in combination with anti-cGMP 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. For example, Sildenafil (Viagra) is an inhibitor of cGMP-specific PDE V, which enhances the vasodilatory effects of cGMP in the corpus cavernosum, and is used to treat erectile dysfunction.

Platform


Fluorescence microplate reader

Excitation540 nm
Emission590 nm
Cutoff570 nm
Recommended plateSolid black

Example protocol


AT A GLANCE

Important      Following protocol only provides a guideline, and should be modified according to your specific needs.

PREPARATION OF STOCK SOLUTIONS

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.

TAMRA-cGMP PDE V stock solution (1 mM)
Make a 1 mM stock solution by adding 500 µL of DMSO into the vial of 0.5 umol TAMRA-cGMP PDE V substrate.

PREPARATION OF WORKING SOLUTION

TAMRA-Cyclic-3’, 5’-GMP PDE V substrate assay solution (2X)
Make 2X TAMRA-Cyclic-3’, 5’-GMP PDE V substrate assay solution by diluting 1 mM TAMRA-Cyclic-3’ ,5’-GMP PDE V 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

  1. Mix equal volume of the PDE V standards or samples with 2X TAMRA-Cyclic-3’ ,5’-GMP PDE V substrate assay solution, and incubate at room temperature for at least 1 hour.
  2. Monitor the fluorescence polarization at Ex/Em = 540/590 nm. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

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


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Correction Factor (260 nm)Correction Factor (280 nm)
FAM-cGMP PDE V substrate *Green Fluorescence*493517830000.320.178

Images


Citations


View all 1 citations: Citation Explorer

References


View all 34 references: Citation Explorer
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
Expression of cAMP and cGMP-phosphodiesterase isoenzymes 3, 4, and 5 in the human clitoris: immunohistochemical and molecular biology study
Authors: Oelke M, Hedlund P, Albrecht K, Ellinghaus P, Stief CG, Jonas U, Andersson KE, Uckert S.
Journal: Urology (2006): 1111
Immunohistochemical distribution of cAMP- and cGMP-Phosphodiesterase (PDE) isoenzymes in the human prostate
Authors: Sampaio FJ., undefined
Journal: Int Braz J Urol (2006): 368
Scanning peptide array analyses identify overlapping binding sites for the signalling scaffold proteins, beta-arrestin and RACK1, in cAMP-specific phosphodiesterase PDE4D5
Authors: Bolger GB, Baillie GS, Li X, Lynch MJ, Herzyk P, Mohamed A, Mitchell LH, McCahill A, Hundsrucker C, Klussmann E, Adams DR, Houslay MD.
Journal: Biochem J (2006): 23
cAMP phosphodiesterase-4A1 (PDE4A1) has provided the paradigm for the intracellular targeting of phosphodiesterases, a process that underpins compartmentalized cAMP signalling
Authors: Huston E, Houslay TM, Baillie GS, Houslay MD.
Journal: Biochem Soc Trans (2006): 504
Expression and Activity of cAMP Phosphodiesterase Isoforms in Pulmonary Artery Smooth Muscle Cells from Patients with Pulmonary Hypertension: Role for PDE1
Authors: Murray F, Patel HH, Suda RY, Zhang S, Thistlethwaite P, Yuan JX, Insel PA.
Journal: Am J Physiol Lung Cell Mol Physiol. (2006)
Immunohistochemical distribution of cAMP- and cGMP-phosphodiesterase (PDE) isoenzymes in the human prostate
Authors: Uckert S, Oelke M, Stief CG, Andersson KE, Jonas U, Hedlund P.
Journal: Eur Urol (2006): 740
The Dunce cAMP phosphodiesterase PDE-4 negatively regulates G alpha(s)-dependent and G alpha(s)-independent cAMP pools in the Caenorhabditis elegans synaptic signaling network
Authors: Charlie NK, Thomure AM, Schade MA, Miller KG.
Journal: Genetics (2006): 111
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
Polymorphisms of the phosphodiesterase 4D, cAMP-specific (PDE4D) gene and risk of ischemic stroke: a prospective, nested case-control evaluation
Authors: Zee RY, Brophy VH, Cheng S, Hegener HH, Erlich HA, Ridker PM.
Journal: Stroke (2006): 2012