ProductDescription
ActivehumanPAI-1functionalassayELISAkit
Plasminogenactivatorinhibitortype1(PAI-1)isinvolvedintheregulationofthebloodfibrinolyticsystem.IncreasedplasmalevelsofPAI-1areimplicatedintheimpairmentoffibrinolyticfunctionandmaybeassociatedwiththromboticdiseases.LevelsofPAI-1increasewithageandareelevatedinconditionssuchasnormalpregnancyandsepsis.ThesensitivequantitativemeasurementoffunctionallyactivehumanPAI-1inplasmasamplesiseasilyperformedwiththis96wellstripformatELISAkit.ThePAI-1concentrationofnormalplatelet-freeplasmais21ng/ml,platelet-richplasmais283ng/mlandserumis270ng/ml.1PAI-1unit=1.34ng.TheassaymeasuresactivePAI-1inthe0.125-100U/mlrange.SamplesgivinghumanPAI-1levelsabove100U/mlshouldbedilutedinPAI-1depletedplasmabeforeuse.Normalplasmashouldbeapplieddirectlytotheplateforbestresults.ItisimportanttoensureaplateletfreepreparationofplasmaasplateletscanreleasePAI-1.FunctionallyactivePAI-1reactswithurokinasecoatedontoamicrotiterplate.LatentorcomplexedPAI-1willnotbindtotheplateandwillnotbedetectedbytheassay.Validatedincitrate,EDTA,andheparinplasma.VitronectindoesnotinterferewiththedetectionofactivePAI-1.ThiskitdoesnotcrossreactwithmousePAI-1.Afterappropriatewashingsteps,antihumanPAI-1primaryantibodybindstothePAI-1.Excessantibodyiswashedaway,andboundantibodyisreactedwiththesecondaryantibodyconjugatedtoHRP.Followinganadditionalwashingstep,TMBsubstrateisusedforcolordevelopmentat450nm.ColordevelopmentisproportionaltotheconcentrationofPAI-1inthesamples.AstandardcalibrationcurveispreparedinPAI-1depletedplasmausingdilutionsofpurifiedPAI-1andismeasuredalongwiththetestsamples.AllreagentsandstandardsareprovidedintheseELISAkits.TheHumanPAI-1standardprovidedinthiskitiscalibratedagainsttheWHOInternationalStandardforHumanPAI-1(NIBSCCode92/654).Thisassayusesanexclusiveenzymecapturetechnologytoonlydetectfunctionallyactiveprotein.
ThestandardcurveforthiskitisgeneratedusingincludedPAI-1depletedhumanplasmaandisbestsuitedformeasurementofPAI-1inplasmaorserum.ThiskitisalsoavailablewithastandardcurveinBSA/TBSforresearchersmeasuringPAI-1inculturemediaortissueextractsundercatalognumberHPAIKT-NP.
Suggestedadditionalreagents:10XWashBuffer,TMBSubstrate,uPAPlate,SecondaryAntibody
GeneID:5054
Swiss-Prot/UniProtID:P05121
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