Description:ShortDescription:InthisassaytheALBUMINpresentinsamplesreactswiththeanti-ALBUMINantibodieswhichhavebeenadsorbedtothesurfaceofpolystyrenemicrotitrewells.Aftertheremovalofunboundproteinsbywashing,anti-AlbuminantibodiesconjugatedwithhorserADIshperoxidase(HRP),areadded.Theseenzyme-labeledantibodiesformcomplexeswiththepreviouslyboundAlbumin.Followinganotherwashingstep,theenzymeboundtotheimmunosorbentisassayedbytheadditionofachromogenicsubstrate,3,3’,5,5’-tetramethylbenzidine(TMB).ThequantityofboundenzymevariesdirectlywiththeconcentrationofAlbumininthesampletested;thus,theabsorbance,at450nm,isameasureoftheconcentrationofAlbumininthetestsample.ThequantityofAlbumininthetestsamplecanbeinterpolatedfromthestandardcurveconstructedfromthestandards,andcorrectedforsampledilution.AdditionalInformation:NameRatAlbuminELISAKitRelatedProductNamesRatAlbuminELISAKit;N/ARatAlbuminALBGeneSymbolALBGeneFullnamealbuminGeneid24186AliasSymbolsAlbumin,ALBProteinNameSerumalbuminDescriptionofTargetSerumalbumin,themainproteinofplasma,hasagoodbindingcapacityforwater,Ca2+,Na+,K+,fattyacids,hormones,bilirubinanddrugs.Itsmainfunctionistheregulationofthecolloidalosmoticpressureofblood.Majorzinctransporterinplasma,SwissprotIDP02770SpeciesReactivityRatAssayInfoAssayMethodology:QuantitativeSandwichELISASensitivity1.620ng/mlKitRange6.25ng/ml-400ng/mlHomologyRatNCBIAccNumberNP_000468.1StandardRangeReferToManualSampleTypeSerum,plasmaandotherBIOLOGicalfluidsProductFormatInSolutionSpecimen/VolumesReferToManualSwissProtNumberP02768ApplicationsELISAReconstitutionandStorage1.DILUENTThe5XDiluentConcentrateisstableuntiltheexpira-tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionsshouldbestoredat4-8C.2.WASHSOLUTIONThe20XWashSolutionConcentrateisstableuntiltheexpira¬tiondate.The1Xworkingsolutionisstableforatleastoneweekfromthedateofpreparation.Bothsolutionscanbestoredatroomtemperature(1625C)orat4-8C.3.ENZYME-ANTIBODYCONJUGATEUndilutedhorseradishperoxidaseanti-Albuminconjugateshouldbestoredat4-8Canddilutedimmediatelypriortouse.Theworkingconjugatesolutionisstableforupto8hours.4.CHROMOGEN-SUBSTRATESOLUTIONTheSubstrateSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.5.STOPSOLUTIONTheStopSolutionshouldbestoredat4-8Candisstableuntiltheexpirationdate.6.ANTI-RATALBUMINELISAMICROPLATEAnti-RatAlbumincoatedwellsarestableuntiltheexpirationdate,andshouldbestoredat4-8Cinsealedfoilpouchwithdesiccantpack.7.RATALBUMINCALIBRATORThelyophilizedRatAlbumincalibratorshouldbestoredat4Corfrozenuntilreconstituted.Thereconstitutedcalibratorshouldbealiquotedoutandstoredfrozen(Avoidmultiplefreeze-thawcycles).Theworkingstandardsolutionsshouldbepreparedimmediatelypriortouseandarestableforupto8hours.ReactivityRatKitSize96wellKitPrincipleGenWayBiotechAlbuminELISAKit(Rat)(GWB-1B2B4B)isbasedonstandardsandwichenzyme-linkedimmuno-sorbentassaytechnology.AnantibodyforSerumalbuminhasbeenpre-coatedontoa96-wellplate(12x8WellStrips).Standardsortestsamplesareaddedtothewells,incubatedandwashedtoremoveunboundproteins.AnAnti-Serumalbumin-HRPconjugateddetectorantibodyisthenadded,incubatedandunboundconjugateiswashedaway.AnenzymaticreactionisproducedthroughtheadditionofTMBsubstratewhichiscatalyzedbyHRPgeneratingabluecolorproductthatchangestoyellowafteraddingacidicstopsolution.Thedensityofyellowcolorationreadbyabsorbanceat450nmisquantitativelyproportionaltotheamountofsampleSerumalbumincapturedinthewell.KitComponentComponentAmountAnti-RatAlbuminMicroplate96Wells(12x8WellStrips)20XDiluentConcentrate50mL20XWashSolution50mL100XEnzyme-AntibodyConjugate150uLChromogen-SubstrateSolution12mLStopSolution12mLRatAlbuminCalibrator1KitDuration70MinutesStorageRefertokitinstructionsforcomponentstorageLeadTimeDomestic:within1weekdelivery International:1weekIntendedUseResearchUseOnlyAdditionalInformationIntendedUse:TheAlbumintestkitsareahighlysensitivetwo-siteenzymelinkedimmunoassay(ELISA)formeasuringalbumininbiologicalsamplesofrats.::Reagents:1.DILUENTCONCENTRATE(RunningBuffer)Onebottlecontaining50mlofa5Xconcentrateddiluentrunningbuffer.2.WASHSOLUTIONCONCENTRATEOnebottlecontaining50mlofa20Xconcentratedwashsolution.3.ENZYME-ANTIBODYCONJUGATE100XOnevialcontaining150ulofaffinitypurifiedanti-RatAlbuminantibodyconjugatedwithhorseradishperoxidaseinastABIlizingbuffer.4.CHROMOGEN-SUBSTRATESOLUTIONOnevialcontaining12mLof3,3’,5,5’-tetramethybenzidine(TMB)andhydrogenperoxideincitricacidbufferatpH3.3.5.STOPSOLUTIONOnevialcontaining12ml0.3Msulfuricacid.WARNING:Avoidcontactwithskin.6.ANTI-RATALBUMINELISAMICROPLATETwelveremovableeight(8)wellmicrowellstripsinwellholderframe.Eachwelliscoatedwithaffinitypurifiedanti-RatAlbumin.7.RATALBUMINCALIBRATOROnevialcontainingalyophilizedRatAlbumincalibrator.