ProductBackground:WhenPIP3isgeneratedviaphosphorylationofPI(4,5)P2(PIP2)byPI3-Kinase,theproductsoftheenzymaticreactioncompetewithbiotinylatedI(1,3,4,5)P4fortheinteractionwiththePIP3detectorprotein.Theinteractionofbiotinylated-I(1,3,4,5)P4andthePIP3detectorproteinbringsbothAcceptorandDonorbeadstogether,producingacascadeofchemicalreactionsandleADIngtotheAlphaScreen®signal.Intheabsenceofthisinteraction,proximityoftheDonorandAcceptorbeadsisdecreased,producingalossofluminescentsignalwhichisinverselyrelatedtoenzymeactivity.AlphaScreen®technologyisbasedontheemissionoflight(520-620nm)byAcceptorbeadsactivatedbytheproximityofDonorbeads.
Thingstoconsider:ThisproductisintendedforusewiththeAmplifiedLuminescentProximityHomogenousAssay(AlphaScreen®)technologyandrequirestheresearchertoobtaindonorandacceptorbeadsfromtheAlphaScreen®GST(Glutathione-S-Transferase)DetectionKit,whichmustbepurchasedseparatelyfromPerkinelmerLifeSciences.AmicroplatereaderwithAlphaScreen®capABIlitiesisalsorequired.
FeaturedinPublications:
1.Drees,B.E.,A.Weipert,etal.(2003)."Competitivefluorescencepolarizationassaysforthedetectionofphosphoinositidekinaseandphosphataseactivity."CombChemHighThroughputScreen6(4):321-330.
2.Boldyreff,B.,T.L.Rasmussen,etal.(2008)."ExpressionandPurificationofPI3Kinase{alpha}andDevelopmentofanATPDepletionandanAlphaScreenPI3KinaseActivityAssay."JBiomolScreen.
3.D'Angelo,N.D.,T.S.Kim,etal.(2011)."Discoveryandoptimizationofaseriesofbenzothiazolephosphoinositide3-kinase(PI3K)/mammaliantargetofrapamycin(mTOR)dualinhibitors."JMedChem54(6):1789-1811.
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ClassIPI3KActivityAssays:Echelonhasmultipleformatsavailable.PleaseseethePI3-KAssayComparisonChart formoreinformation.
ClassIIIPI3KActivityAssay(K-3000)
LipidMassELISA's:PIP3MassELISA(K-2500s)andPI(4,5)P2MassELISA(K-4500)
HumanPI3KEnzymes:alpha(p110α/p85α),beta(p110β/p85alphα),PI3KαααE545K–ActiveMutant(p110α/p85α)
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OptimizedPI3Kenzymebuffer(K-KBZ)
NoticetoPurchaser:EchelonBiosciencesproductsaresoldforresearchanddevelopmentpurposesonlyandarenottobeincorporatedintoproductsforresalewithoutwrittenpermissionfromEchelonBiosciences.Thiskitandallnon-radioactive,competitiveassaysfordeterminingphosphoinositide-3-kinase(PI3-K)activityarecoveredbyEchelonBiosciencesInc.U.S.Patent7,067,269.Thepurchaseofthisproductincludesalimited,non-transferableimmunityfromsuitundertheforegoingpatentclaimsforusingonlythisamountofproductforthepurchaser'sowninternalresearch.
ProductKeywords:PI3-Kinase,PI3K,Phosphatidylinositol3-kinases,PI3-kinases,PI3Ks,K-1000,PIP3,PI(345)P3,PtdIns(3,4,5)P3,Phosphatidylinositol(3,4,5)-triphosphate
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