CPF4

FRET sensor molecule for detection of PDE activity



ALX-620-079-C250	250 µg	364.00 USD

ALX-620-079-C500	500 µg	670.00 USD

Enzyme-cleavable FRET sensor molecule for the ratiometric measurement of phosphodiesterase activity. This is an effective method for the monitoring of phosphodiesterase activity in real time with high sensitivity; a large spectral shift in the emission spectrum is noticed after hydrolysis of the phosphodiester linker by phosphodiesterase I (Ex(max): 370nm; Em(max) uncleaved: 515nm; Em(max) cleaved: 450nm). The role of NPPs (pyrophosphatases/phosphodiesterases) can also be investigated.

Product Specification

Alternative Name:	Coumarin phosphodiester-linked fluorescein

Formula:	C₄₄H₂₈ClN₂O₁₄P

MW:	875.1

Purity:	≥99% (¹H-NMR)

Appearance:	Yellow solid.

Solubility:	Soluble in DMSO (10mM) or in aqueous buffers, pH>8.5.

Long Term Storage:	+4°C

Handling:	Keep cool and dry. Protect from light.

Background / Technical Information:	FRET (fluorescence resonance energy transfer) is a radiationless transmission of an energy quantum between two fluorophores which are in close proximity. Cleavage of a chemical bond between these fluorophores, which have an overlap in their spectra, results in a large shift in the emission spectra. Addition of phosphodiesterase I, which catalyzes the hydrolysis of phosphodiester bonds (substrates are nucleic acids and cyclic nucleotides), to an aqueous solution of CPF4 results in an increase in the donor fluorescence and a decrease in the acceptor fluorescence. CPF4 is an effective tool for the monitoring of the activity of phosphodiesterases in real time with high sensitivity.

Figure: Sensor principle.

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Product Literature References

Design and synthesis of an enzyme-cleavable sensor molecule for phosphodiesterase activity based on fluorescence resonance energy transfer: H. Takakusa, et al.; J. Am. Chem. Soc. 124, 1653 (2002), Abstract;

Design and Synthesis of Intramolecular Resonance-Energy Transfer Probes for Use in Ratiometric Measurements in Aqueous Solution: Y. Kawanishi, et al.; Angew. Chem. Int. Ed. Engl. 39, 3438 (2000), Abstract;