Description:ShortDescription:InthisassaytheMyeloperoxidasepresentinserumsamplereactswiththeanti-MPOantibodieswhichhavebeenadsorbedtothesurfaceofpolystyrenemicrotitrewells.Aftertheremovalofunboundserumproteinsbywashing,anti-MPOantibodiesconjugatedwithhorserADIshperoxidase(HRP),areadded.Theseenzyme-labeledantibodiesformcomplexeswiththepreviouslyboundserumMPO.Followinganotherwashingstep,theenzymeboundtotheimmunosorbentisassayedbytheadditionofachromogenicsubstrate,3,3’,5,5’-tetramethylbenzidine(TMB).ThequantityofboundenzymevariesdirectlywiththeconcentrationofMyeloperoxidaseinthesampletested;thus,theabsorbance,at450nm,isameasureoftheconcentrationofMyeloperoxidaseinthetestsample.ThequantityofMyeloperoxidaseinthetestsamplecanbeinterpolatedfromthestandardcurveconstructedfromthestandards,andcorrectedforserumdilution.AdditionalInformation:NameHumanMyeloperoxidaseELISAKitRelatedProductNamesHumanMyeloperoxidaseELISAKit;MPO;EC1.11.1.7HumanMyeloperoxidaseMPOGeneSymbolMPOGeneFullnamemyeloperoxidaseGeneid4353AliasSymbolsMPOProteinNameMyeloperoxidaseDescriptionofTargetPartofthehostdefensesystemofpolymorphonuclearleukocytes.ItisresponsIBLeformicrobicidalactivityagainstawiderangeoforganisms.InthestimulatedPMN,MPOcatalyzestheproductionofhypohalousacids,primarilyhypochlorousacidinphysiolSwissprotIDP05164SpeciesReactivityHumanAssayInfoAssayMethodology:QuantitativeSandwichELISASensitivity0.994ng/mlKitRange1.875ng/ml-120ng/mlHomologyHumanNCBIAccNumberNP_000241.1StandardRangeReferToManualSampleTypeSerum,plasmaandotherBIOLOGicalfluidsProductFormatInSolutionSpecimen/VolumesReferToManualSwissProtNumberP05164ApplicationsELISAReconstitutionandStorage1.DILUENTThe5XDiluentConcentrateisstableuntiltheexpira-tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionsshouldbestoredat4-8C.2.WASHSOLUTIONThe20XWashSolutionConcentrateisstableuntiltheexpira¬tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionscanbestoredatroomtemperature(1625C)orat4-8C.3.ENZYME-ANTIBODYCONJUGATEUndilutedhorseradishperoxidaseanti-MPOconjugateshouldbestoredat4-8Canddilutedimmediatelypriortouse.Theworkingconjugatesolutionisstableforupto8hours.4.CHROMOGEN-SUBSTRATESOLUTIONTheSubstrateSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.5.STOPSOLUTIONTheStopSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.6.ANTI-HUMANMYELOPEROXIDASEELISAMICROPLATEAnti-HumanMPOcoatedwellsarestableuntiltheexpirationdate,andshouldbestoredat4-8Cinsealedfoilpouchwithdesiccantpack.7.HUMANMYELOPEROXIDASECALIBRATORThelyophilizedHumanMyeloperoxidasecalibratorshouldbestoredat4Corfrozenuntilreconstituted.Thereconstitutedcalibratorshouldbealiquotedoutandstoredfrozen(Avoidmultiplefreeze-thawcycles).Theworkingstandardsolutionsshouldbepreparedimmediatelypriortouseandarestableforupto1hours.ReactivityHumanKitSize96wellKitPrincipleGenWayBiotechMyeloperoxidaseELISAKit(Human)(GWB-2795F0)isbasedonstandardsandwichenzyme-linkedimmuno-sorbentassaytechnology.AnantibodyforMyeloperoxidasehasbeenpre-coatedontoa96-wellplate(12x8WellStrips).Standardsortestsamplesareaddedtothewells,incubatedandwashedtoremoveunboundproteins.AnAnti-Myeloperoxidase-HRPconjugateddetectorantibodyisthenadded,incubatedandunboundconjugateiswashedaway.AnenzymaticreactionisproducedthroughtheadditionofTMBsubstratewhichiscatalyzedbyHRPgeneratingabluecolorproductthatchangestoyellowafteraddingacidicstopsolution.Thedensityofyellowcolorationreadbyabsorbanceat450nmisquantitativelyproportionaltotheamountofsampleMyeloperoxidasecapturedinthewell.KitComponentComponentAmountAnti-HumanMyeloperoxidaseMicroplate96Wells(12x8WellStrips)20XDiluentConcentrate50mL20XWashSolution50mL100XEnzyme-AntibodyConjugate150uLChromogen-SubstrateSolution12mLStopSolution12mLHumanMyeloperoxidaseCalibrator1KitDuration90MinutesStorageRefertokitinstructionsforcomponentstorageLeadTimeDomestic:within1weekdelivery International:1weekIntendedUseResearchUseOnlyAdditionalInformationIntendedUse:TheMyeloperoxidasetestkitsareahighlysensitivetwo-siteenzymelinkedimmunoassay(ELISA)formeasuringMyeloperoxidaseinserum,plasmaofHuman.::Reagents:1.DILUENTCONCENTRATE(RunningBuffer)Onebottlecontaining50mlofa5Xconcentrateddiluentrunningbuffer.2.WASHSOLUTIONCONCENTRATEOnebottlecontaining50mlofa20Xconcentratedwashsolution.3.ENZYME-ANTIBODYCONJUGATE100XOnevialcontaining150ulofaffinitypurifiedanti-HumanMyeloperoxidaseantibodyconjugatedwithhorseradishperoxidaseinastABIlizingbuffer.4.CHROMOGEN-SUBSTRATESOLUTIONOnevialcontaining12mLof3,3’,5,5’-tetramethybenzidine(TMB)andhydrogenperoxideincitricacidbufferatpH3.3.5.STOPSOLUTIONOnevialcontaining12ml0.3Msulfuricacid.WARNING:Avoidcontactwithskin.6.ANTI-HUMANMYELOPEROXIDASEELISAMICROPLATETwelveremovableeight(8)wellmicrowellstripsinwellholderframe.Eachwelliscoatedwithaffinitypurifiedanti-HumanMPO.7.HUMANMYELOPEROXIDASECALIBRATOROnevialcontainingalyophilizedHumanMyeloperoxidasecalibrator.