雌酮（E1）检测试剂盒说明书

文档简介雌酮(E1)检测试剂盒适用生物General，通用雌酮(E1)检测试剂盒检测范围24.69-2000pg/mL灵敏度8.86pg/mL样本类型Serum,plasmaandotherbiologicalfluids.实验时长2.5h实验方法竞争YZ法雌酮(E1)检测试剂盒规格96T雌酮(E1)检测试剂盒ELISAKitforEstrone(E1)FORINVITROANDRESEARCHUSEONLY,NOTFORUSEINCLINICALDIAGNOSTICPROCEDURES!OrganismspeciesGeneralProductNo.CEB003GeSampletypeSerum,plasmaandotherbiologicalfluids.Format96-wellstripplateAssaylength2.5hoursDetectionrange24.69-2000pg/mLThestandardcurveconcentrationsusedfortheELISA’swere2000pg/mL,666.67pg/mL,222.22pg/mL,74.07pg/mL,24.69pg/mLSensitivityTheminimumdetectabledoseofthiskitistypicallylessthan8.86pg/mL.SpecificityThisassayhashighsensitivityandexcellentspecificityfordetectionofEstrone(E1).Nosignificantcross-reactivityorinterferencebetweenEstrone(E1)andanalogueswasobserved.RecoveryMatriceslistedbelowwerespikedwithcertainlevelofrecombinantEstrone(E1)andtherecoveryrateswerecalculatedbycomparingthemeasuredvaluetotheexpectedamountofEstrone(E1)insamples.MatrixRecoveryrange(%)Average(%)serum(n=5)80-9085EDTAplasma(n=5)97-105102heparinplasma(n=5)96-105101PrecisionIntra-assayPrecision(Precisionwithinanassay):3sampleswithlow,middleandhighlevelEstrone(E1)weretested20timesononeplate,respectively.Inter-assayPrecision(Precisionbetweenassays):3sampleswithlow,middleandhighlevelEstrone(E1)weretestedon3differentplates,8replicatesineachplate.CV(%)=SD/meanX100Intra-Assay:CV<10%Inter-Assay:CV<12%LinearityThelinearityofthekitwasassayedbytestingsamplesspikedwithappropriateconcentrationofEstrone(E1)andtheirserialdilutions.Theresultsweredemonstratedbythepercentageofcalculatedconcentrationtotheexpected.Sample1:21:41:81:16serum(n=5)93-101%84-98%84-98%87-96%EDTAplasma(n=5)92-105%93-101%79-90%91-99%heparinplasma(n=5)88-101%91-102%83-99%93-101%StabilityThestabilityofkitisdeterminedbythelossrateofactivity.Thelossrateofthiskitislessthan5%withintheexpirationdateunderappropriatestoragecondition.Tominimizeextrainfluenceontheperformance,operationproceduresandlabconditions,especiallyroomtemperature,airhumidity,incubatortemperature首ldbestrictlycontrolled.Itisalsostronglysuggestedthatthewholeassayisperformedbythesameoperatorfromthebeginningtotheend.ReagentsandmaterialsprovidedReagentsQuantityReagentsQuantityPre-coated,readytouse96-wellstripplate1Platesealerfor96wells4Standard2StandardDiluent1×20mLDetectionReagentA1AssayDiluentA1×12mLDetectionReagentB1×120μLAssayDiluentB1×12mLReagentDiluent1×300μLStopSolution1×6mLTMBSubstrate1×9mLInstructionmanual1WashBuffer(30×concentrate)1×20mLAssayproceduresummary1.Prepareallreagents,samplesandstandards;2.Add50μLstandardorsampletoeachwell.Andthenadd50μLpreparedDetectionReagentAimmediately.Shakeandmix.Incubate1hourat37oC;3.Aspirateandwash3times;4.Add100μLpreparedDetectionReagentB.Incubate30minutesat37oC;5.Aspirateandwash5times;6.Add90μLSubstrateSolution.Incubate15-25minutesat37oC;7.Add50μLStopSolution.Readat450nmimmediately.TestprincipleThisassayemploysthecompetitiveinhibitionenzymeimmunoassaytechnique.AmonoclonalantibodyspecifictoEstrone(E1)hasbeenpre-coatedontoamicroplate.AcompetitiveinhibitionreactionislaunchedbetweenbiotinlabeledEstrone(E1)andunlabeledEstrone(E1)(Standardsorsamples)withthepre-coatedantibodyspecifictoEstrone(E1).Afterincubationtheunboundconjugateiswashedoff.Next,avidinconjugatedtoHorseradishPeroxidase(HRP)isaddedtoeachmicroplatewellandincubated.TheamountofboundHRPconjugateisreverseproportionaltotheconcentrationofEstrone(E1)inthesample.Afteradditionofthesubstratesolution,theintensityofcolordevelopedisreverseproportionaltotheconcentrationofEstrone(E1)inthesample.