Cyclicnucleotidesarehydrolyzedandcompartmentalizedbyafamilyofenzymescalledphosphodieterases.Inmammalsatleast11differentfamiliesofPDEscanbediscriminated(PDE1-PDE11)basedontheirkineticpropertiesandinhibitiontovariouspharmacologicalagents.ThemembersofthePDE3familiesareknownascGMPinhibitedPDEs.ThePDE3enzymesbindbothcGMPandcAMPwithdifferentaffinities(cAMP4-10-foldthancGMP).Thephosphodiesterasetype-3(PDE3)familyiscomprisedof2genes(PDE3AandPDE3B).PDE3Aisa145kDproteinandisexpressedinvarioustissues(myocardium,vascularandnonvascularsmoothmuscles,megakaryocytesandasubsetofneurons(Snyder1999;Maeccietal.,1992).PDE3Bis1112aalongandhastwosplicevariantswhichareexpressedinADIpocytetissue,hepatocyte,kidney,spermatocytesandembryonicneuroepithelium(Reinhatdtet.al.,1995).BothPDE3AandPDE3BaretherapeutictargetsforthetreatmentofobesityasinhibitionofPDE3increasethelipolyticactivityandaltertheinsulinsecretionanditsresponse.PDE3inhibitorsalsopromoteThermogenicandcardiotoniceffectsinanimals.||Source:|RecombinanthumanPDE3B,aa592-end,withN-terminalGSTtag,expressedinanBaculovirusinfectedSf9cellexpressionsystem.||MolecularWeight:86kD||SpecificActivity:|Lot-specific.Samplelotvalue:1270pmol/min/ug||UnitDefinition:|1unitisdefinedastheamountofenzymethatwillconvert1pmoleof3',5'-cAMPto5'AMPperminat30°Cinareactionbuffer.||AssayConditions:|10mMTris-HCl,pH7.4,10mMMgCl2,1mMMnCl2,200uMcAMP,2.5kU5'nucleotidase,37°C,20min.Quantifiedby5'-nucleotidasecleavingthe5'-AMPproductandreleasingthephosphategroupwhichisdetectedbyMalachiteGreenReagent.||Application:|Usefulforthestudyofenzymekinetics,screeninginhibitors,andselectivityprofiling.Otherapplicationsnottested.||RecommendedDilution:|Optimaldilutiondeterminedbytheresearcher.||StorageandStABIlity:|Storeat-70°C.Stableforatleast6months.Avoidfreeze-thawcycles.ormaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.

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