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5'-Nucleotidase (from Crotalus atrox venom)

 
BML-KI307-5000 5000 kU 168.00 USD
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Product Specification

Quantity:5000kU in 1000µl
 
Formulation:Liquid. In TRIS-HCl, pH 7.4, 125mM sodium chloride, 1mM magnesium chloride and 10% (v/v) glycerol.
 
Activity:One U will release 1pmol phosphate per minute from 200µM 5’-AMP, 30°C in a reaction buffer of 10mM Tris-HCl, pH 7.4, 0.2mM MgCl2.
 
Application Notes:5’-nucleotidase is useful as the second enzyme in coupled assays of phosphodiesterases (3’,5’-cyclic nucleotide phosphodiesterase; EC 3.1.4.17). Phosphodiesterase catalyzed hydrolysis of, for example, cAMP to 5’-AMP is coupled to 5’-nucleotidase cleavage of 5’-AMP to inorganic phosphate and adenosine. Phosphodiesterase activity is then quantified by colorimetric detection of the inorganic phosphate with a reagent such as BIOMOL Green (Prod. No. BML-AK111; see also Cyclic Nucleotide Phosphodiesterase Assay Kit, Prod. No. BML-AK800).
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Use/Stability:Stable for at least 6 freeze-thaw cycles, but care should be taken that both freezing and thawing steps are done quickly (e.g. liquid N2 or dry ice/EtOH for freezing, rapid finger-rubbing for thawing). Frozen storage of smaller aliquots is recommended if more than 6 freeze-thaws of the entire vial contents (1000µl) are anticipated. Thawed enzyme is stable on ice for at least 2 hrs.
 
Scientific Background:5’-nucleotidase (5’-ribonucleotide phosphohydrolase; EC 3.1.3.5) is partially purified from Crotalus atrox (Western Diamondback Rattlesnake) venom and is free of detectable phosphodiesterase activity.
 
Protocol:5000 kU is sufficient for 100 coupled phosphodiesterase assays (50'000 U/10µl per well). Coupled reactions carried out in a final reaction volume of 50µl with 0.25mU phosphodiesterase (1 mU = 1nmol 3’,5’-cAMP to 5’-AMP per minute at pH 7.4, 30°C) in 10mM Tris-HCl, pH 7.4, 0.2mM MgCl2, 200µM 3’,5’-cAMP, 30°C. See protocol for Cyclic Nucleotide Phosphodiesterase Assay Kit, Prod. No. BML-AK800, for assay details.
 

Product Literature References

Single inhibition of either PDE3 or PDE4 unmasks β2-adrenoceptor-mediated inotropic and lusitropic effects in the left but not right ventricular myocardium of rat: F. Soler, et al.; Eur. J. Pharmacol. 765, 429 (2015), Abstract;

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