孔雀石绿检测卡（水样）

孔雀石绿检测卡（水样）

本文由北京华安麦科生物技术有限公司整理汇编

2012-08-24 00:00 306阅读次数

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malachitegreen（MG）ELISA1UsepurposeThiskitforFeed,aquaticsamples,watersamplesintheMGremainingquantitativedetection.2ExperimentalprincipleThiskitadoptsmethodsincompetitionELISAInmicroplatescoatedwithantigenconjugatedMG,addMGstandardorsamples,FreeMGandpre-coatedonstripsofMGconjugatedantigencompeteagainstMGantibodyconjugates,WithTMBchromogenicsubstrate,thecolorischangedfrombluetoyellowafteraddingstopsolution,enzymestandardinstrument450nmwavelengthsintesting,absorblightvalueandinthesamplewasMGcontentisinverselyproportionaltothestandardcurve,throughcalculationsampleswasMGconcentrations.3Materialsprovidedwiththekit3.1Microelisastripplate:1block（12well×8strips）.3.2MGstandard:sixvialsof（1ml/vial）,contentisrespectively:0PPB,0.1PPB,0.3PPBand0.9PPB,2.7PPB,8.1PPB.3.3Anti-MGantibodyconjugate:1vial（6ml）.3.4ChromogenSolutionA:1vial（6ml）.3.5ChromogenSolutionB:1vial（6ml）.3.6StopSolution:1vial（6ml）,2Msulphuricacid.3.7sampledilution:1vial（10x,6ml）,usedforsampledilutedwith.3.8washsolution:1vial（20x,20ml）,usedforwashingboard.3.9Instruction.4Neednotprovidematerials4.1equipment4.4.1wavelength450nmmicroplate.4.1.2shredder.4.1.3LiangTong.4.1.4oscillators.4.1.5funnel.4.1.6WhatmanNo1orequivalentfilterpaper.4.1.7traceremoveliquiddevice.4.2reagent4.2.1thedeionizedwaterordistilledwater.4.2.2methanol.5Storage5.1kitstoredin2~8℃,Don’tfrozen5.2Don’tuseuptheMicroelisastripplate首ldbesealeddryingpreserve6.Precautions6.1pleasereadtheinstructionscarefully,beforeuseingthekit.6.2don'tuseexpiredkit.6.3beforeusingthekit,pleaseletthereagentrecovertoroomtemperature（25+2℃）,theproposalforatleast2hourstotemperature.6.4thestandardcontainMG,payspecialattentionto,theoperation,we首ldbringgloves.6.5stopsolutioniscontainingsulfate,whenusing,preventburnsskinandcorrosionclothing.6.6differentstandardandsamplesuctionheadusedcannotbemixeduse,otherwise,itwillaffecttestresults.6.7differentbatchesofreagentkitnotmix,Differentstandardandsamplesuctionheadshallnotbeusedincombination,otherwise,itwillaffecttheexperimentalresult.6.8dilutedsamplemustusethiskitofsamplediluent,otherwise,itwillaffecttheexperimentalresult6.9mixedreagents首ldavoidblistering.7Workingliquidpreparation7.1Carbendazimstandards:0ppb,0.1PPB,0.3PPBand0.9PPB,2.7PPB,8.1PPB7.2washsolution:1:20withdistilledwater（1+19）diluted.preparation7.3sampledilutions:1:10withdistilledwater（1+9）diluted.preparation7.3ChromogenSolutionreagent:alreadystandby,avoidlightstraightas7.4stopsolution:alreadyterminationaside8Sampleprocessingprogram（sampleinextractionprocess,muststrictlyaccordingtotheoperationoftheextractionprocess首ldbeaccuratedilution,canappearotherwiseresultsareinaccurate,samples首ldbekeptinacoolplacetoavoidlightandfrozenkeep）8.1Smashthesamplestaken10g,add20ml70%methanolsolution8.2powerfuloscillation3minutes8.3WhatmanNo1filterwith8.4Takethe25μltreatmentofthesamplebyadding25μlsampledilutioninthewells（sampledilutionfactorof2）9Enzyme2lindedanalysissteps9.1experimentalguidelines9.1.1experimentbeginspriortoallreagentinboxesoutsidetheroomtemperature（25fullyrecoveredto+2℃）,timeabout2hours.Returntoroomtemperature（25+2℃）againafterremoveStrips,excessporebartosealimmediatelytothe2~8℃dryingpreserveNote:besuretotemperature,otherwisefullyguaranteetheaccuracyandprecisionoftheaffectingdetection.9.1.2afterusepleaseimmediatelyreagentputback2~8℃preservation9.1.3pleasedon'tchangeanalysisprogram9.1.4pleaseuseaccuratetraceremoveliquiddevice9.1.5operationoncestarted,pleasedonotinterruptanyprogram9.1.6ELISAresultsofrepeatabilityofseveredependsonoperatingprocedures,pleasestrictlyaccordingtorequirementsoperation9.1.7toavoidcross-contamination,eachstandardandsamples首ldusedifferentsuctionheadaddsamples9.1.8plussampledomakesuckaheadcontactmicroporousthesolutionorinsidesurface9.2analysissteps9.2.1beforehandnumbered,markB0,standardandthesamplepositionrecommendeddoubleorificedetection9.2.2taketherequiredamountoftheStrips（Stripsdetachable）,willspareattribwattleandsealedimmediatelyputbackagain2~8℃preservation9.2.3sampledilutions（10）,washsolutionx（20x）dilutionintoworkingliquid（distilledwaterordeionizedwaterdilute）In9.2.4B0welljoining50μl0.0ng/mlstandard9.2.5ineachstandardwelljoining50μlstandard9.2.6ineachsamplewelljoining50μlsamplesolution9.2.7Inallwelljoining50μlAnti-MGantibodyconjugate9.2.8gentlysloshingresponseboardforafewseconds.9.337℃warmbath30min（warmbathprocesssometimespatreactionplate,canreducedoubleorificeerror）9.3.1UncoverClosureplatemembrane,discardLiquid,drybyswing,addwashingbuffertoeverywell,stillfor30sthendrain,repeat5times,drybypat..9.4reaction9.4.1washingprocedurecompleted,immediatelywithliquidapparatusineverytracemovemicroporousfirstjoin50μlChromogenSolutionA,add50μlChromogenSolutionB,Slightsloshingresponseboardmakethoroughlyincorporated379.4.2℃warmbath10min9.4.3eachwelljoining50μlStopSolution,blending9.4.4in450nmtestingabsorbency,resultin5mininsideread.10Theresultscalculated10.1quantitativeanalysis10.1.1obtainedbyeachconcentrationstandardsolutionandtheaveragevalueofasamplespectrophotometry（B）dividedbythefirststandard（0standardabsorbencyvalue（B0）multiplyingby1**%,namelypercentageabsorbencyvalues.B-standardsolutionorsamplesolutionofaverageabsorbanceofthevaluesB0-0μg/Lstandardsolutionofaverageabsorbanceofthevalues10.1.2withMGconcentrationsofvaluesfortheXaxis,100centabsorbanceofthevalueoftheYaxis,drawstandardcurve.Accordingtothesamplepercentageabsorbencyvalues,whichgetcorrespondingpointsfromcurve,namelytheabscissadenotesthemulti-goalMGconcentrationonthenumericalcurvature.theagainstseveralnamelytodetermineMGconcentrationC（ppb）10.1.3becausethesampleafterdilutedinadvance,soaccordingtothestandardcurvegainsfromdifferentconcentrationsamplesmustagainmultiplythedilutedtimes.10.2halfquantitatively10.1.1visualhalfquantitativedetermination:first,chooseanappropriatestandardfluidandsampleswithoperation,accordingtothesamplesandstandardsubstanceabsorbencyvaluethediscretionofthejudgeiscompared,samplechromavalueislessthanorgreaterthanstandardvalues.10.1.2instrumenthalfquantitativedetermination:first,chooseanappropriatestandardfluidandsampleswithoperation,accordingtothesampleandstandardcolordepthcomparison,judgesamplechromavalueislessthanorgreaterthanstandardvalues.11SpecificityPhysicalcrossreactionMG1**%CrystalViolet:95%Recessivemalachitegreen:0.1%HiddenCrystalViolet:<0.1%12KitparametersThiskitdetectionlimitis0.05PPBB0absorbanceoftheoptimalvalue首ldbegreaterthan1.0Kitabsorbencyboardinsideerrorislessthan8%,boardbetweenerrorislessthan15%.Withthismanualprovidedatissuesampleextractionmethodrecoveryisgreaterthan80%.13AnalysisrestrictionThiskittestingpositiveforsamples首lduseanothermethodsuchasHPLCorGC/MStobeverified.上海恒远生物科技有限公司，专业销售“孔雀石绿试剂盒”品质保证，值得信赖！如果你想了解该产品的详细信息，欢迎来电来函！[详细]
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