ThePI3-KBetaInhibitorScreeningServiceisacompetitiveassayfortheproductofthePI3-Kinasereaction,PI(3,4,5)P₃.Theenzymeusedintheserviceispurifiedrecombinanthumanp110β/p85αactiveenzymeandthesubstrateisdiC₈PI(4,5)P₂.DuplicatePI3Kreactionsareperformedinthepresenceofeachsamplecompoundprovidedbythecustomer.AfterthePI3Kreactionsarecomplete,reactionproductsaredetectedusingLuminescentAssayforPI3-KinaseActivity(AlphaScreen®Assay,cat#K-1300).

ThefullPI3-KBetaInhibitorScreeningServiceisdividedintotwoparts–(Stage1)screeningforpotentialinhibitortargetinghumanPI3Kp110β/p85α,and(Stage2)determinationofIC50valueforhitcompoundfromstage1.Whentestingforinhibitionpotentialinstage1,asingleselectedcompoundconcentration(determinedbycustomer,usuallyahighdose)forallcompoundsampleswillbetested.Assaycontrolsinclude“PositiveReactioncontrol”withoutcompoundorinhibitor;“PositiveInhibitioncontrols”withawellknownPI3Kinhibitor;“Substratecontrol”withoutenzymeandinhibitor;and“CompoundInterferencecontrol”withoutenzyme.API(3,4,5)P₃Standardcurvecanbeincludeduponrequestwithextracharge.Whenasamplecompoundshowsinhibitionpotential(ahit)andfurthercharacterizationisrequired,IC50valueofthecompoundcanbedeterminedthroughcompoundtitrationastestservicestage2.

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