Description:ShortDescription:InthisassaytheIgEpresentinserumsamplereactswiththeanti-IgEantibodieswhichhavebeenadsorbedtothesurfaceofpolystyrenemicrotitrewells.Aftertheremovalofunboundserumproteinsbywashing,anti-IgEantibodiesconjugatedwithhorserADIshperoxidase(HRP),areadded.Theseenzyme-labeledantibodiesformcomplexeswiththepreviouslyboundserumIgE.Followinganotherwashingstep,theenzymeboundtotheimmunosorbentisassayedbytheadditionofachromogenicsubstrate,3,3’,5,5’-tetramethylbenzidine(TMB).ThequantityofboundenzymevariesdirectlywiththeconcentrationofIgEinthesampletested;thus,theabsorbance,at450nm,isameasureoftheconcentrationofIgEinthetestsample.ThequantityofIgEinthetestsamplecanbeinterpolatedfromthestandardcurveconstructedfromthestandards,andcorrectedforserumdilution.AdditionalInformation:NameHumanIgEELISAKitRelatedProductNamesHumanIgEELISAKit;N/AHumanIgEIGHEAliasSymbolsIGHE,IgepsilonchainCregion,IgEProteinNameImmunoglobulinheavyconstantepsilonDescriptionofTargetConstantregionofimmunoglobulinheavychains.Immunoglobulins,alsoknownasantibodies,aremembrane-boundorsecretedglycoproteinsproducedbyBlymphocytes.Intherecognitionphaseofhumoralimmunity,themembrane-boundimmunoglobulinsserveasreSwissprotIDP01854SpeciesReactivityHumanAssayInfoAssayMethodology:QuantitativeSandwichELISASensitivity0.912ng/mlKitRange3.125ng/ml-200ng/mlHomologyHumanStandardRangeReferToManualSampleTypeSerum,plasmaandotherBIOLOGicalfluidsProductFormatInSolutionSpecimen/VolumesReferToManualSwissProtNumberP01854ApplicationsELISAReconstitutionandStorage1.DILUENTThe5XDiluentConcentrateisstableuntiltheexpira-tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionsshouldbestoredat4-8C.2.WASHSOLUTIONThe20XWashSolutionConcentrateisstableuntiltheexpira¬tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionscanbestoredatroomtemperature(1625C)orat4-8C.3.ENZYME-ANTIBODYCONJUGATEUndilutedhorseradishperoxidaseanti-IgEconjugateshouldbestoredat4-8Canddilutedimmediatelypriortouse.Theworkingconjugatesolutionisstableforupto8hours.4.CHROMOGEN-SUBSTRATESOLUTIONTheSubstrateSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.5.STOPSOLUTIONTheStopSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.6.ANTI-HUMANIgEELISAMICROPLATEAnti-HumanIgEcoatedwellsarestableuntiltheexpirationdate,andshouldbestoredat4-8Cinsealedfoilpouchwithdesiccantpack.7.HUMANIgECALIBRATORLongTermStorage:Uponreceipt,aliquotthecalibratorandstorethemfrozen.Theywillbestableuntilexpirationdate.ShortTermStorage:thecalibratorisstableforupto14daysat4C.Theworkingstandardsolutionsshouldbepreparedimmediatelypriortouseandarestableforupto8hours.ReactivityHumanKitSize96wellKitPrincipleGenWayBiotechIgEELISAKit(Human)(GWB-2E2ECD)isbasedonstandardsandwichenzyme-linkedimmuno-sorbentassaytechnology.AnantibodyforImmunoglobulinheavyconstantepsilonhasbeenpre-coatedontoa96-wellplate(12x8WellStrips).Standardsortestsamplesareaddedtothewells,incubatedandwashedtoremoveunboundproteins.AnAnti-Immunoglobulinheavyconstantepsilon-HRPconjugateddetectorantibodyisthenadded,incubatedandunboundconjugateiswashedaway.AnenzymaticreactionisproducedthroughtheadditionofTMBsubstratewhichiscatalyzedbyHRPgeneratingabluecolorproductthatchangestoyellowafteraddingacidicstopsolution.Thedensityofyellowcolorationreadbyabsorbanceat450nmisquantitativelyproportionaltotheamountofsampleImmunoglobulinheavyconstantepsiloncapturedinthewell.KitComponentComponentAmountAnti-HumanIgEMicroplate96Wells(12x8WellStrips)20XDiluentConcentrate50mL20XWashSolution50mL100XEnzyme-AntibodyConjugate150uLChromogen-SubstrateSolution12mLStopSolution12mLHumanIgECalibrator1KitDuration80MinutesStorageRefertokitinstructionsforcomponentstorageLeadTimeDomestic:within1weekdelivery International:1weekIntendedUseResearchUseOnlyAdditionalInformationIntendedUse:ThetotalHumanIgEtestkitisahighlysensitivetwo-siteenzymelinkedimmunoassay(ELISA)formeasuringIgEinhumanbiologicalsamples.::Reagents:1.DILUENTCONCENTRATE(RunningBuffer)Onebottlecontaining50mlofa5Xconcentrateddiluentrunningbuffer.2.WASHSOLUTIONCONCENTRATEOnebottlecontaining50mlofa20Xconcentratedwashsolution.3.ENZYME-ANTIBODYCONJUGATE100XOnevialcontaining150ulofaffinitypurifiedanti-HumanIgEantibodyconjugatedwithhorseradishperoxidaseinastABIlizingbuffer.4.CHROMOGEN-SUBSTRATESOLUTIONOnevialcontaining12mLof3,3’,5,5’-tetramethybenzidine(TMB)andhydrogenperoxideincitricacidbufferatpH3.3.5.STOPSOLUTIONOnevialcontaining12ml0.3Msulfuricacid.WARNING:Avoidcontactwithskin.6.ANTI-HUMANIgEELISAMICROPLATETwelveremovableeight(8)wellmicrowellstripsinwellholderframe.Eachwelliscoatedwithaffinitypurifiedanti-HumanIgE.7.HUMANIgECALIBRATOROnevialcontainingofHumanIgEcalibrator.