logo
AAT Bioquest

FAM-cAMP PDE IV substrate *Green Fluorescence*

This green fluorescent 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.

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. FAM-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 FAM-cAMP PDE IV substrate.

PREPARATION OF WORKING SOLUTION

FAM-Cyclic-3’, 5’-AMP PDE IV substrate assay solution (2X):
Make 2X FAM-Cyclic-3’, 5’-AMP PDE IV substrate assay solution by diluting 1 mM FAM-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

  1. Mix equal volume of the PDE IV standards or samples with 2X FAM-Cyclic-3’ ,5’-AMP PDE IV substrate assay solution, and incubate at room temperature for at least 1 hour.

  2. Monitor the fluorescence polarization at Ex/Em = 490/525 nm.

Spectrum

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Correction Factor (260 nm)Correction Factor (280 nm)
TAMRA-cAMP PDE IV substrate *Red Fluorescence*552578900000.320.178

References

View all 34 references: Citation Explorer
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
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)
Page updated on December 11, 2024

Ordering information

Price
Unit size
Catalog Number13602
Quantity
Add to cart

Additional ordering information

Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
Technical SupportContact us
Purchase orderSend to sales@aatbio.com
ShippingStandard overnight for United States, inquire for international
Request quotation

Physical properties

Molecular weight

~800

Solvent

DMSO

Spectral properties

Correction Factor (260 nm)

0.32

Correction Factor (280 nm)

0.178

Extinction coefficient (cm -1 M -1)

83000

Excitation (nm)

493

Emission (nm)

517

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

Platform

Fluorescence microplate reader

Excitation490 nm
Emission525 nm
Cutoff515 nm
Recommended plateSolid black
Product Image
Product Image
Gallery Image 1