髓过氧化物酶（MPO）检测试剂盒说明书

深度解析

髓过氧化物酶(MPO)检测试剂盒

检测范围0.781-50ng/mL灵敏度0.35ng/mL
样本类型Serum,plasma,tissuehomogenates,celllysates,cellculturesupernatesandotherbiologicalfluids.
实验时长4.5h实验方法双抗夹心法

髓过氧化物酶(MPO)检测试剂盒ELISAKitforMyeloperoxidase(MPO)
FORINVITROANDRESEARCHUSEONLY,NOTFORUSEINCLINICALDIAGNOSTICPROCEDURES!

OrganismspeciesHomosapiens(Human)
ProductNo.SEA601Hu
SampletypeSerum,plasma,tissuehomogenates,celllysates,cellculturesupernatesandotherbiologicalfluids.
Format96-wellstripplate
Assaylength4.5hours
Detectionrange0.781-50ng/mLThestandardcurveconcentrationsusedfortheELISA’swere50ng/mL,25ng/mL,12.5ng/mL,6.25ng/mL,3.125ng/mL,1.563ng/mL,0.781ng/mL
SensitivityTheminimumdetectabledoseofthiskitistypicallylessthan0.35ng/mL.

髓过氧化物酶(MPO)检测试剂盒Specificity
ThisassayhashighsensitivityandexcellentspecificityfordetectionofMyeloperoxidase(MPO).
Nosignificantcross-reactivityorinterferencebetweenMyeloperoxidase(MPO)andanalogueswasobserved.
Recovery
MatriceslistedbelowwerespikedwithcertainlevelofrecombinantMyeloperoxidase(MPO)andtherecoveryrateswerecalculatedbycomparingthemeasuredvaluetotheexpectedamountofMyeloperoxidase(MPO)insamples.
髓过氧化物酶(MPO)检测试剂盒

MatrixRecoveryrange(%)Average(%)
serum(n=5)95-10298
EDTAplasma(n=5)79-9185
heparinplasma(n=5)81-8984

Precision
Intra-assayPrecision(Precisionwithinanassay):3sampleswithlow,middleandhighlevelMyeloperoxidase(MPO)weretested20timesononeplate,respectively.
Inter-assayPrecision(Precisionbetweenassays):3sampleswithlow,middleandhighlevelMyeloperoxidase(MPO)weretestedon3differentplates,8replicatesineachplate.
CV(%)=SD/meanX100
Intra-Assay:CV<10%
Inter-Assay:CV<12%
Linearity
ThelinearityofthekitwasassayedbytestingsamplesspikedwithappropriateconcentrationofMyeloperoxidase(MPO)andtheirserialdilutions.Theresultsweredemonstratedbythepercentageofcalculatedconcentrationtotheexpected.
髓过氧化物酶(MPO)检测试剂盒

Sample1:21:41:81:16
serum(n=5)98-105%87-97%87-104%88-105%
EDTAplasma(n=5)90-98%94-101%85-104%91-98%
heparinplasma(n=5)98-105%85-97%93-101%91-99%

Stability
Thestabilityofkitisdeterminedbythelossrateofactivity.Thelossrateofthiskitislessthan5%withintheexpirationdateunderappropriatestoragecondition.
Tominimizeextrainfluenceontheperformance,operationproceduresandlabconditions,especiallyroomtemperature,airhumidity,incubatortemperature首ldbestrictlycontrolled.Itisalsostronglysuggestedthatthewholeassayisperformedbythesameoperatorfromthebeginningtotheend.
Reagentsandmaterialsprovided

ReagentsQuantityReagentsQuantity
Pre-coated,readytouse96-wellstripplate1Platesealerfor96wells4
Standard2StandardDiluent1×20mL
DetectionReagentA1×120μLAssayDiluentA1×12mL
DetectionReagentB1×120μLAssayDiluentB1×12mL
TMBSubstrate1×9mLStopSolution1×6mL
WashBuffer(30×concentrate)1×20mLInstructionmanual1

Assayproceduresummary
1.Prepareallreagents,samplesandstandards;
2.Add100μLstandardorsampletoeachwell.Incubate2hoursat37oC;
3.Aspirateandadd100μLpreparedDetectionReagentA.Incubate1hourat37oC;
4.Aspirateandwash3times;
5.Add100μLpreparedDetectionReagentB.Incubate30minutesat37oC;
6.Aspirateandwash5times;
7.Add90μLSubstrateSolution.Incubate15-25minutesat37oC;
8.Add50μLStopSolution.Readat450nmimmediately.
髓过氧化物酶(MPO)检测试剂盒Testprinciple
ThetestprincipleappliedinthiskitisSandwichenzymeimmunoassay.Themicrotiterplateprovidedinthiskithasbeenpre-coatedwithanantibodyspecifictoMyeloperoxidase(MPO).Standardsorsamplesarethenaddedtotheappropriatemicrotiterplatewellswithabiotin-conjugatedantibodyspecifictoMyeloperoxidase(MPO).Next,AvidinconjugatedtoHorseradishPeroxidase(HRP)isaddedtoeachmicroplatewellandincubated.AfterTMBsubstratesolutionisadded,onlythosewellsthatcontainMyeloperoxidase(MPO),biotin-conjugatedantibodyandenzyme-conjugatedAvidinwillexhibitachangeincolor.Theenzyme-substratereactionisterminatedbytheadditionofsulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm±10nm.TheconcentrationofMyeloperoxidase(MPO)inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.

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2004-09-12