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美國GeneScience 厭氧工作站AG300,A30,A20,Anaerobox IV型厭氧培養(yǎng)箱
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大鼠 IV型膠原 (Co IV) ELISA 檢測試劑盒
- 試驗(yàn)原理:CoIV試劑盒是固相夾心法酶聯(lián)免疫吸附實(shí)驗(yàn)(ELISA).已知CoIV濃度的標(biāo)準(zhǔn)品、未知濃度的樣品加入微孔酶標(biāo)板內(nèi)進(jìn)行檢測。先將CoIV和生物素標(biāo)記的抗體同時(shí)溫育。洗滌后,加入親和素標(biāo)記過的HRP。再經(jīng)過溫育和洗滌,去除未結(jié)合的酶結(jié)合物,然后加入底物A、B,和酶結(jié)合物同時(shí)作用。產(chǎn)生顏色。顏色的深淺和樣品中CoIV的濃度呈比例關(guān)系。試劑盒內(nèi)容及其配制試劑盒成份(2-8℃保存)96孔配置48孔配置配制96/48人份酶標(biāo)板1塊板(96T)半塊板(48T)即用型塑料膜板蓋1塊半塊即用型標(biāo)準(zhǔn)品:800ng/ml1瓶(0.6ml)1瓶(0.3ml)按說明書進(jìn)行稀稀空白對照1瓶(1.0ml)1瓶(0.5ml)即用型標(biāo)準(zhǔn)品稀釋緩沖液1瓶(5ml)1瓶(2.5ml)即用型生物素標(biāo)記的抗CoIV抗體1瓶(6ml)1瓶(3.0ml)即用型親和鏈酶素-HRP1瓶(10ml)1瓶(5.0ml)即用型洗滌緩沖液1瓶(20ml)1瓶(10ml)按說明書進(jìn)行稀釋底物A1瓶(6.0ml)1瓶(3.0ml)即用型底物B1瓶(6.0ml)1瓶(3.0ml)即用型終止液1瓶(6.0ml)1瓶(3.0ml)即用型標(biāo)本稀釋液1瓶(12ml)1瓶(6.0ml)即用型自備材料蒸餾水。加樣器:5ul、10ul、50ul、100ul、200ul、500ul、1000ul。振蕩器及磁力攪拌器等。安全性避免直接接觸終止液和底物A、B。一旦接觸到這些液體,請盡快用水沖洗。實(shí)驗(yàn)中不要吃喝、抽煙或使用化妝品。不要用嘴吸取試劑盒里的任何成份。[詳細(xì)]
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2018-09-19 10:01
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厭氧培養(yǎng)箱
- 厭氧培養(yǎng)箱[詳細(xì)]
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2012-10-31 00:00
安裝說明
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BINDER CO2/O2培養(yǎng)箱厭氧方面的應(yīng)用
- BINDER CO2/O2培養(yǎng)箱厭氧方面的應(yīng)用[詳細(xì)]
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2024-09-28 00:42
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大鼠(Rat)IV型膠原(Co IV)ELISA試劑盒英文說明書
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Normal07.8磅02falsefalsefalseMicrosoftInternetExplorer4Goatanti-RatCollagenTypeIVCollectSampleserumorbloodplasmaStorage:2-8°CPackagesize:96determinationsPRINCIPLEOFTHEMETHODTheCoIVkitisasolidphasephasesandwichenzymelinkedimmunosorbentassay(ELISA).Samples,includingstandardsofknownCoIVconcentrationsandunknownsarepipettedintothesewells.Duringthefirstincubation,theCoIVantigenandabiotinylatedmonoclonalantibodyspecificforCoIVaresimultaneouslyincubated.Afterwashing,theenzyme(streptavidin-peroxydase)isadded.Afterincubationandwashingtoremovealltheunboundenzyme,asubstratesolutionwhichisactingontheboundenzymeisaddedtoinduceacolouredreactionproduct.TheintensityofthiscolouredproductisdirectlyproportionaltotheconcentrationofCoIVpresentinthesamples.REAGENTSPROVIDEDANDRECONSTTTUTIONREAGENTS(Storeat2-8℃)1×96WELLS0.5×96WELLSRECONSTTTUTION96/48-wellsmicrotiterplates10.5Ready-to-usePlastivcover21Ready-to-useStandard:800ng/ml1Vials(0.6ml)0.5Vials(0.3ml)Seereagentspreparationonpage3Blankcontrol1Vials(1.0ml)1Vials(0.5ml)Ready-to-useStandardDiluent1Vials(5ml)1Vials(2.5ml)Ready-to-useBiotinylatedanti-CoIV1Vials(6ml)1Vials(3.0ml)Ready-to-useStreptavidin-HRP1Vials(10ml)1Vials(5.0ml)Ready-to-useWashingBuffer1Vials(20ml)1Vials(10ml)50×concentrateSubstrateA1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSubstrateB1Vials(6.0ml)1Vials(3.0ml)Ready-to-useStoppingSolution1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSampleDiluent1Vials(12ml)1Vials(6.0ml)Ready-to-useMATERIALREQUIREDBUTNOTPROVIDEDlDistilledwaterlPipettes:10ul、50ul、100ul、200ul、1000ul。lVortexmixerandmagneticstirrer.SAFETYlForresearchuseonlylAvoidanyskincontactwithH2SO4andTMB.Incaseofcontact,washthoroughlywater.lDonoteat,drink,smokeorapplycosmeticswherekitreagentsareused.lDonotpipettebymouth.PROCEDURALNOTES/LAB.QUALITYCONTROLlWhennotinuse,kitcomponents首ldbestoredrefrigeratedorfrozenasindicatedonvialsorbottles.Allreagents首ldbewarmedtoroomtemperaturebeforeuse.Lyophilizedstandards首ldbediscardedafteruse.lOncethedesirednumberofstripshasbeenremoved,immediatelyresealthebagtoprotecttheremainingstripsfromedterioration.lCoverorcapallreagentswhennotinuse.lDonotmisorinterchangereagentsbetweendifferentlots.lDonotusereagentsbeyondtheexpirationdateofthekit.lUseacleandisposableplasticpipettetipforeachreagent,standard,orspecimenadditioninordertoavoidcross-contamination,forthedispensingofH2SO4andsubstratesolution,avoidpipetteswithmetalparts.lUseacleanplasticcontainertopreparethewashingsolution.lThoroughlymixthereagentsandsamplesbeforeusebyagitationorswir領(lǐng).lAllresidualwashingliquidmustbedrainedfromthewellsbyefficientaspirationorbydecantationfollowedbytappingtheplateforcefullyonabsorbentpaper.Neverinsertabsorbentpaperdirectlyintothewells.lTheTMBsolutionislightsensitive.Avoidprolongedexposuretolight,also,avoidcontactoftheTMBsolutionwithmetaltopreventcolourdevelopment.WarningTMBistoxicavoiddirectcontactwithhands.Disposeoffproperly.Ifadarkbluecolourdevelopswithinafewminutesafterpreparation,thisindicatesthattheTMBsolutionhasbeencontaminatedandmustbediscarede.Readabsorbanceswithin1houraftercompletionoftheassay.lWhenpipettingreagents,maintainaconsistentorderofadditionfromwell-to-well.Thiswillensureequalincubationtimesforallwells.lRespectincubationtimesdescribedintheassayprocedure.SPECIMENCOLLECTION\PROCESSINGANDSTORAGElSerum---Avoidanyinintentionalstimulationofthecellsbytheprocedure.Usepyrogen\endotoxinfreecollectingtubes.Serum首ldberemovedrapidlyandcarefullyfromtheredcellsafterclothing.Forthat,afterclothing,centrifugeatapproximately1000×gfor10minandremoveserum.lPlasma---EDTA\citrateandheparinplasmacanbeassayed.Spinsamplesat1000×gfor30minremoveparticulates.Harvestplasma.lCellculturesupernatants---Removeparticulatesandaggregatesbyspinningatapproximately1000×gfor10min.lStorage---Ifnotanalyzedshortlyaftercollection,samples首ldbealiquoted(250-500ul)toavoidfreeze-thawcyclesandstoredfrozenat-70℃.Avoidmultiplefreeze-thawcyclesoffrozenspecimens.Whenpossible,avoiduseofbadlyhemolyzedorlipemicsera.Iflargeamountsofparticlesarepresent,this首ldberemovedpriortoassaybycentrifugationorfiltration.lRecommendation---Donotthawbyheatingat37℃or56℃.Thawatroomtemperatureandmakesurethatsampleiscompletelythawedandhomogenousbeforeassaying.PREPARATIONOFREAGENTSlStandards:Standardhavetobereconstituledwiththevolumeofstandardbufferdiluentindicatedonthevial.Thisreconstitutionproducesastocksolutionof800ng/mlCoIV.Allowstandardtostandfor5lminuteswithgentleswir領(lǐng)priortomakingdilutions.Serialdilutionsofstandardmustbemadebeforeeachassysandcannotbestored.800ng/ml(6Standard)Originaldensity50ul。400ng/ml(5Standard)100ul6Standard+100uldiludent200ng/ml(4Standard)100ul5Standard+100uldiludent100ng/ml(3Standard)100ul4Standard+100uldiludent50ng/ml(2Standard)100ul3Standard+100uldiludent25ng/ml(1Standard)100ul2Standard+100uldiludent0ng/mlBlankControl50ul。lWashingbuffer50×concentrate:Dilute50timesindistilledwater.ASSAYMETHODlBeforeuse,mixallreagentsthoroughlywithoutmakingfoam.lDeterminethenumberofmicrowellstripsrequiredtotestthedesirednumberofsamples,plusappropriatenumberofwellsneededforrunningblanksstandards.Eachsample,standardandblank首ldbeassayedinduplicate.Removesufficientmicrowellstripsfromthepouch.lAdd50ulofstandarddiluenttostandardwellsB1,B2,C1,C2,D1,D2,E1,E2,F1,F2.Reconstitutestandardvialwiththeappropriatevolumeasdescribedinthechapterreagentspreparation.Preparation.Pipet100ulofstandardintowellsA1andA2(seeplateschemebelow).Transfer50ulfromA1andA2toB1andB2wells.Mixthecontentsbyrepeatedaspirationsandejections.Takecarenottoscratchtheinnersurfaceofmicrowells.RepatthisprocedurefromthewellsB1,B2towellsC1,C2andfromwellsC1,C2toD1,D2andsooncreatingtwoparallelrowsofCoIVstandarddilutionsranging,Add50ulofstandarddiluenttotheblandwells.lDilutesamples1:1distribing50ulofsampleinto50ulofdilluent,Add50ulofdilutedsampletowells..lAdd50ulofdilutedbiotinylatedanti-CoIVtoallwells.lCoverwithaplatevoverandincubatefor1hourat37℃.lRemovethecoverandwashtheplateasfollows:⑴ aspiratetheliquidfromeachwell,⑵ dispensse0.3mlofwashingsolutionintoeachwell.⑶ Aspirateagainthecontetofeachwellafter0.5minute.⑷ Repeatsteps⑵and⑶threetimes.lDistribute80ulofstreptavidin-HRPsolutiontoallwells,includingblankwells.lCoverandincubate30minat37℃.lRemovethecoverandemptywells,Washmicrowellstripsaccordingtostep,Proceedimmediatelytothenextstep.lAdd50ulSubstrateAandSubstrateBtoeachwell。Incubatefor10minat37℃。lTheenzyme-substratereactionisstoppedbyquicklypipetting50ulofH2SO4.stopreagentintoeachwell,includingtheblankwells,tocompletelyanduniformlyinactivatetheenzyme.ResultsmustberedimmediatelyaftertheadditionofH2SO4.lReadabsorbanceofeachwellonaspectrophotometerusing450nmastheprimarywavelengthandoptionally620nm(610nmto650nmisacceptable)asthereferencewavelength.SUGGESTEDPLATESCHEMEStandardconcentrations(ng/ml)A800800samplesamplesamplesamplesamplesamplesamplesamplesamplesampleB400400samplesamplesamplesamplesamplesamplesamplesamplesamplesampleC200200samplesamplesamplesamplesamplesamplesamplesamplesamplesampleD100100samplesamplesamplesamplesamplesamplesamplesamplesamplesampleE5050samplesamplesamplesamplesamplesamplesamplesamplesamplesampleF2525samplesamplesamplesamplesamplesamplesamplesamplesamplesampleG00samplesamplesamplesamplesamplesamplesamplesamplesamplesampleHsamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesampleLIMITATIONSOFTHEPROCEDUREDonotextrapolatethestandardcurvebeyondthemaxstandardcurvepoint.Thedose-responseisnon-linearinthisregionandgoodaccruacyisdifficulttoobtain.CALCULATIONOFRESULTSTheminimumdetectableconcentrationinthisassayisestimatedtobe1.0ng/ml[詳細(xì)]
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2018-11-16 10:02
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