ProductBackground:TheassayisacompetitiveELISAinwhichthesignalisinverselyproportionaltotheamountofPIP3detected.OncePIP3hasbeenextractedfrommillionsofcellsamples,itisfirstincubatedwithaPIP3detectorprotein,thenaddedtothePIP3-coatedmicroplateforcompetitivebinding.Aperoxidase-linkedsecondarydetectionreagentandcolorimetricsubstrateisusedtodetectthePIP3detectorproteinboundtotheplate.ThecolorimetricsignalisinverselyproportionaltotheamountofPIP3extractedfromcells.
TheproductionofPIP3fromPI(4,5)P2(PIP2)byclassIPI3-kinases(PI3-K)isimportantinmultiplecellsignalingpathways.Typically,experimentstomeasurePI3-Kactivityhaveinvolvedphosphorylationofaphosphoinositidesubstrateusing32P,thenextractionofrADIoactiveproducts,andseparationusingthin-layerchromatography.TheassayplatemethoddevelopedbyEchelonBiosciences,Inc.allowstheusertodeterminePI3-KactivitybymeasuringtheamountofPIP3extractedfromcellsbymeansofstandardELISAformat,eliminatingtheneedforradioactivity,andthinlayerchromatography.
FeaturedinPublications:
1.Ehrmantrout,KimberlyK.,Branch,AngieM.,Neilsen,PaulO.,andThompson,JamesE.."NonradioactiveQuantificationofPhosphoinositide3-Kinase(PI3K)ProductsfromBreastCancerCellLines".AmericanSocietyforCellBIOLOGy(ASCB),Washington,DC.2007.
2.Ehrmantrout,KimberlyK.,Branch,AngieM.,Neilsen,PaulO.."QuantificationofPI3-KinaseProductsfromBreastCancerCellLines".AmericanAssociationforCancerResearch(AACR).Denver,CO.2009.
3.King,JS.,Teo,R.,etal.“TheMoodStABIlizerLithiumSuppressesPIP3SignalinginDictyosteliumandHumanCells.”DiseaseModelsandMechanisms2(5-6):306-12
4.Beneteau,M.,Pizon,M.,etal.“LocalizationogFas/CD95intotheLipidRaftsonDown-ModulationofthePhosphatidylinositol3-KinaseSignalingPathway.”MolCancerRes6(4):604-613
5.Costa,C.,Ebi,H.,etal."MeasurementofPIP3LevelsRevealsanUnexpectedRoleforp110betainEarlyAdaptiveResponsestop110alpha-SpecificInhibitorsinLuminalBreastCancer"Cell27(1):97-108
6.Gross,C.,C.-W.Chang,etal.(2015)."IncreasedExpressionofthePI3KEnhancerPIKEMediatesDeficitsinSynapticPlasticityandBehaviorinFragileXSyndrome."CellReports,DOI:10.1016/j.celrep.2015.03.060
7.S.Choi,etal."AgoNIST-stimulatedphosphatidylinositol-3,4,5-trisphosphategenerationbyscaffoldedphosphoinositidekinases"NatureCellBiology(2016)18(12),1324-35,doi:10.1038/ncb3441
RelatedProducts:
Echelon'sPIPMassAssays
PI(4,5)P2MassELISA
PI(3,4)P2MassELISA
PI3Kinase:PI3Kalphaenzyme,active,PI3KInhibitors,andPI3KActivityELISA
ProteinKinasesandPhosphatases
Keywords:Phosphatidylinositol(3,4,5)-trisphosphate,PtdIns(3,4,5)P3,PI(3,4,5)P3,PIP3,Phosphatidylinositol3-kinase,PI3-kinase,PI3-Kinase,PI3-K,PI3K,PI3Kinase,PI3K,cellextraction,lipidextraction |
