ProductDescription
ActivehumanuPAfunctionalassayELISAkit
Urokinaseplasminogenactivator(uPA),alongwithitsreceptoruPAR,isaserineproteasethatactivatesplasminogentoplasmininthebloodfibrinolyticsystem.Itisalsoimplicatedineventsrelatedtocellinvasionandmigration.ClinicalstudieshaveindicatedthathighuPAlevelsmayelevatetheriskfortumorinvasionandmetastasis.Increasedexpressionandactivitycanexertpotentarthritogenicpropertiesinrheumatoidarthritispatients.IncreaseduPAactivitymaybeanimplicationforthepathophysiologyofendometriosis.ThesensitivequantitativemeasurementoffunctionallyactivehumanuPAinplasmaandotherBIOLOGicalfluidsiseasilyperformedwiththis96wellstripformatELISAkit.ThemeanvalueofuPAinhealthydonorswasfoundtobe1.1ng/ml.TheassaymeasuresactiveuPAinthe0.1-50ng/mlrange.SamplesgivinghumanuPAlevelsabove50ng/mlshouldbedilutedinblockingbufferbeforeuse.SamplesofhumanplasmaincitrateorEDTAmaybeassayedwiththiskit.Plasmainheparinisnotrecommended.ItisimportanttoensureaplateletfreepreparationasplateletscanreleasePAI-1,whichinturncouldpotentiallyformacomplexwithactiveuPA.SerumandcellculturemediaatneutralpHmayalsobeused.FunctionallyactiveuPAwillformacovalentcomplexwiththebiotinylatedhumanPAI-1whichisboundtotheavidinontheplate.InactiveorcomplexeduPAwillnotbindtothePAI-1andwillnotbedetectedbytheassay.ThiskitdoesnotcrossreactwithmouseuPA.Afterappropriatewashingsteps,anti-humanuPAprimaryantibodybindstothecapturedenzyme.Excessantibodyiswashedaway,andboundantibodyisreactedwiththesecondaryantibodyconjugatedtoHRP.Followinganadditionalwashingstep,TMBsubstrateisusedforcolordevelopmentat450nm.ColordevelopmentisproportionaltotheconcentrationofactiveuPAinthesamples.AstandardcalibrationcurveispreparedusingdilutionsofpurifieduPAandismeasuredalongwiththetestsamples.AllreagentsandstandardsareprovidedintheseELISAkits.
Thisassayusesanexclusiveenzymecapturetechnologytoonlydetectfunctionallyactiveprotein.
Suggestedadditionalreagents:10XWashBuffer,TMBSubstrate,AvidinPlate,SecondaryAntibody
GeneID:5328
Swiss-Prot/UniProtID:P00749
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AdditionalInformation
| Applications | |
|---|---|
| Source | Multiple |
| Epitope | Urokinase |
| Host | Human |
| Species | Human |
| Storage | 4C |
| ShippingConditions | Coldpack |
| Expiration | 12monthsfrommanufacture,seelabelforexpirationdate |
| Form | Completekit |
| SampleSize | 1kit |
| GeneName | PLAU |
| GeneID | 5328 |
| UniProtID | P00749 |