Description:ShortDescription:InthisassaytheIgG1presentinserumsamplereactswiththeanti-IgG1antibodieswhichhavebeenadsorbedtothesurfaceofpolystyrenemicrotitrewells.Aftertheremovalofunboundserumproteinsbywashing,anti-IgG1antibodiesconjugatedwithhorserADIshperoxidase(HRP),areadded.Theseenzyme-labeledantibodiesformcomplexeswiththepreviouslyboundserumIgG1.Followinganotherwashingstep,theenzymeboundtotheimmunosorbentisassayedbytheadditionofachromogenicsubstrate,3,3’,5,5’-tetramethylbenzidine(TMB).ThequantityofboundenzymevariesdirectlywiththeconcentrationofIgG1inthesampletested;thus,theabsorbance,at450nm,isameasureoftheconcentrationofIgG1inthetestsample.ThequantityofIgG1inthetestsamplecanbeinterpolatedfromthestandardcurveconstructedfromthestandards,andcorrectedforserumdilution.AdditionalInformation:NameMouseIgG1ELISAKitRelatedProductNamesMouseIgG1ELISAKit;N/AMouseIgG1IGHG1AliasSymbolsIGHG1,Iggamma,ImmunoglobulinG,Iggamma-1chainCregionProteinNameIgG1SpeciesReactivityMouseAssayInfoAssayMethodology:QuantitativeSandwichELISASensitivity0.645ng/mlKitRange6.25ng/ml-200ng/mlHomologyMouseStandardRangeReferToManualSampleTypeSerum,plasmaandotherBIOLOGicalfluidsProductFormatInSolutionSpecimen/VolumesReferToManualSwissProtNumberP01857ApplicationsELISAReconstitutionandStorage1.DILUENTThe5XDiluentConcentrateisstableuntiltheexpira-tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionsshouldbestoredat4-8C.2.WASHSOLUTIONThe20XWashSolutionConcentrateisstableuntiltheexpira¬tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionscanbestoredatroomtemperature(1625C)orat4-8C.3.ENZYME-ANTIBODYCONJUGATEUndilutedhorseradishperoxidaseanti-IgG1conjugateshouldbestoredat4-8Canddilutedimmediatelypriortouse.Theworkingconjugatesolutionisstableforupto8hours.4.CHROMOGEN-SUBSTRATESOLUTIONTheSubstrateSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.5.STOPSOLUTIONTheStopSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.6.ANTI-MOUSEIgG1ELISAMICROPLATEAnti-MouseIgG1coatedwellsarestableuntiltheexpirationdate,andshouldbestoredat4-8Cinsealedfoilpouchwithdesiccantpack.7.MOUSEIgG1CALIBRATORLongTermStorage:Uponreceipt,aliquotthecalibratorandstorethemfrozen.Theywillbestableuntilexpirationdate.ShortTermStorage:thecalibratorisstableforupto14daysat4C.Theworkingstandardsolutionsshouldbepreparedimmediatelypriortouseandarestableforupto8hours.ReactivityMouseKitSize96wellKitPrincipleGenWayBiotechIgG1ELISAKit(Mouse)(GWB-30CDC5)isbasedonstandardsandwichenzyme-linkedimmuno-sorbentassaytechnology.AnantibodyforIgG1hasbeenpre-coatedontoa96-wellplate(12x8WellStrips).Standardsortestsamplesareaddedtothewells,incubatedandwashedtoremoveunboundproteins.AnAnti-IgG1-HRPconjugateddetectorantibodyisthenadded,incubatedandunboundconjugateiswashedaway.AnenzymaticreactionisproducedthroughtheadditionofTMBsubstratewhichiscatalyzedbyHRPgeneratingabluecolorproductthatchangestoyellowafteraddingacidicstopsolution.Thedensityofyellowcolorationreadbyabsorbanceat450nmisquantitativelyproportionaltotheamountofsampleIgG1capturedinthewell.KitComponentComponentAmountAnti-MouseIgG1Microplate96Wells(12x8WellStrips)20XDiluentConcentrate50mL20XWashSolution50mL100XEnzyme-AntibodyConjugate150uLChromogen-SubstrateSolution12mLStopSolution12mLMouseIgG1Calibrator1KitDuration90MinutesStorageRefertokitinstructionsforcomponentstorageLeadTimeDomestic:within1weekdelivery International:1weekIntendedUseResearchUseOnlyAdditionalInformationIntendedUse:TheIgG1testkitsareahighlysensitivetwo-siteenzymelinkedimmunoassay(ELISA)formeasuringIgG1inbiologicalsamplesofmice.::Reagents:1.DILUENTCONCENTRATE(RunningBuffer)Onebottlecontaining50mlofa5Xconcentrateddiluentrunningbuffer.2.WASHSOLUTIONCONCENTRATEOnebottlecontaining50mlofa20Xconcentratedwashsolution.3.ENZYME-ANTIBODYCONJUGATE100XOnevialcontaining150ulofaffinitypurifiedanti-MouseIgG1antibodyconjugatedwithhorseradishperoxidaseinastABIlizingbuffer.4.CHROMOGEN-SUBSTRATESOLUTIONOnevialcontaining12mLof3,3’,5,5’-tetramethybenzidine(TMB)andhydrogenperoxideincitricacidbufferatpH3.3.5.STOPSOLUTIONOnevialcontaining12ml0.3Msulfuricacid.WARNING:Avoidcontactwithskin.6.ANTI-MOUSEIgG1ELISAMICROPLATETwelveremovableeight(8)wellmicrowellstripsinwellholderframe.Eachwelliscoatedwithaffinitypurifiedanti-MouseIgG1.7.MOUSEIgG1CALIBRATOROnevialcontainingaMouseIgG1calibrator.