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Human HMW Adiponectin Quantikine ELISA Kit
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HumanHMWAdiponectinQuantikineELISAKit
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Human HMW Adiponectin Quantikine ELISA Kit- HumanHMWAdiponectinQuantikineELISAKit[詳細]
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2018-10-01 10:01
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- Human uPAR Quantikine ELISA Kit
- HumanuPARQuantikineELISAKit[詳細]
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2018-10-01 10:01
產(chǎn)品樣冊
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- Human VEGF Immunoassay Quantikine ELISA
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QuantikineELISAThispackageinsertmustbereadinitsentiretybeforeusingthisproduct.Forresearchuseonly.Notforuseindiagnosticprocedures.CatalogNumberDVE00CatalogNumberSVE00CatalogNumberPDVE00ForthequantitativedeterminationofhumanVascularEndothelialGrowthFactor（VEGF）concentrationsincellculturesupernates,serum,andplasma.MANUFACTUREDANDDISTRIBUTEDBY:USA&Canada|R&DSystems,Inc.614McKinleyPlaceNE,Minneapolis,MN55413,USATEL:（800）343-7475（612）379-2956FAX:（612）656-4400E-MAIL:info@RnDSystems.comDISTRIBUTEDBY:UK&Europe|R&DSystemsEurope,Ltd.19BartonLane,AbingdonSciencePark,AbingdonOX143NB,UKTEL:+44（0）1235529449FAX:+44（0）1235533420E-MAIL:info@RnDSystems.co.ukChina|R&DSystemsChinaCo.,Ltd.24A1HuaMinEmpirePlaza,726WestYanAnRoad,ShanghaiPRC200050TEL:+86（21）52380373FAX:+86（21）52371001E-MAIL:info@RnDSystemsChina.com.cnTABLEOFCONTENTSSECTIONPAGEINTRODUCTION.....................................................................................................................................................................1PRINCIPLEOFTHEASSAY...................................................................................................................................................2LIMITATIONSOFTHEPROCEDURE.................................................................................................................................2TECHNICALHINTS.................................................................................................................................................................2MATERIALSPROVIDED&STORAGECONDITIONS...................................................................................................3OTHERSUPPLIESREQUIRED.............................................................................................................................................4PRECAUTIONS.........................................................................................................................................................................4SAMPLECOLLECTION&STORAGE.................................................................................................................................4REAGENTPREPARATION.....................................................................................................................................................SSAYPROCEDURE.............................................................................................................................................................6CALCULATIONOFRESULTS...............................................................................................................................................7TYPICALDATA.........................................................................................................................................................................7PRECISION................................................................................................................................................................................8RECOVERY................................................................................................................................................................................8SENSITIVITY.............................................................................................................................................................................8LINEARITY.................................................................................................................................................................................9CALIBRATION..........................................................................................................................................................................9SAMPLEVALUES..................................................................................................................................................................10SPECIFICITY...........................................................................................................................................................................11REFERENCES.........................................................................................................................................................................12PLATELAYOUT.....................................................................................................................................................................13www.RnDSystems.com1INTRODUCTIONVascularendothelialgrowthfactor（VEGForVEGF-A）,alsoknownasvascularpermeabilityfactor（VPF）,isapotentmediatorofbothangiogenesisandvasculogenesisinthefetusandadult（1-3）.ItisamemberofthePDGFfamilythatischaracterizedbythepresenceofeightconservedcysteineresiduesinacystineknotstructureandtheformationofantiparalleldisulfide-linkeddimers（4）.Humansexpressalternatelysplicedisoformsof121,145,165,183,189,and206aminoacids（aa）inlength（4）.VEGF165appearstobethemostabundantandpotentisoform,followedbyVEGF121andVEGF189（3,4）.IsoformsotherthanVEGF121containbasicheparin-bindingregionsandarenotfreelydiffusible（4）.HumanVEGF165shares88%aasequenceidentitywithcorrespondingregionsofmouseandratVEGF.VEGFisexpressedinmultiplecellsandtissuesincludingskeletalandcardiacmuscle（5,6）,hepatocytes（7）,osteoblasts（8）,neutrophils（9）,macrophages（10）,keratinocytes（11）,brownadiposetissue（12）,CD34+stemcells（13）,endothelialcells（14）,fibroblasts,andvascularsmoothmusclecells（15）.VEGFexpressionisinducedbyhypoxiaandcytokinessuchasIL-1,IL-6,IL-8,oncostatinMandTNF-α（3,4,9,16）.VEGFisoformsaredifferentiallyexpressedduringdevelopmentandintheadult（3）.VEGFdimersbindtotworelatedreceptortyrosinekinases,VEGFR1（alsocalledFlt-1）andVEGFR2（Flk-1/KDR）,andinducetheirhomodimerizationandautophosphorylation（3,4,7,17,18）.Thesereceptorshavesevenextracellularimmunoglobulin-likedomainsandanintracellularsplittyrosinekinasedomain.Theyareexpressedonvascularendothelialcellsandarangeofnon-endothelialcells.AlthoughVEGFaffinityishighestforbindingtoVEGFR1,VEGFR2appearstobetheprimarymediatorofVEGFangiogenicactivity（3,4）.VEGF165alsobindsthesemaphorinreceptor,neuropilin-1,whichpromotescomplexformationwithVEGFR2（19）.VEGFisbestknownforitsroleinvasculogenesis.Duringembryogenesis,VEGFregulatestheproliferation,migration,andsurvivalofendothelialcells（3,4）,thusregulatingbloodvesseldensityandsize,butplayingnoroleindeterminingvascularpatterns.VEGFpromotesboneformationthroughosteoblastandchondroblastrecruitmentandisalsoamonocytechemoattractant（20-22）.Afterbirth,VEGFmaintainsendothelialcellintegrityandisapotentmitogenformicro-andmacro-vascularendothelialcells.Inadults,VEGFfunctionsmainlyinwoundhea領(lǐng)andthefemalereproductivecycle（3）.Indiseasedtissues,VEGFpromotesvascularpermeability.Itisthusthoughttocontributetotumormetastasisbypromotingbothextravasationandtumorangiogenesis（23,24）.VariousstrategieshavebeenemployedtherapeuticallytoantagonizeVEGF-mediatedtumorangiogenesis（25）.CirculatingVEGFlevelscorrelatewithdiseaseactivityinautoimmunediseasessuchasrheumatoidarthritis,multiplesclerosisandsystemiclupuserythematosus（26）.TheQuantikineHumanVEGFImmunoassayisa4.5hoursolidphaseELISAdesignedtomeasureVEGF165levelsincellculturesupernates,serum,andplasma.ItcontainsSf21-expressedrecombinanthumanVEGF165andantibodiesraisedagainsttherecombinantprotein.ResultsobtainedfornaturallyoccurringhumanVEGFandrecombinanthumanVEGF121showedlinearcurvesthatwereparalleltothestandardcurvesobtainedusingtheQuantikineHumanVEGFImmunoassaystandards.TheseresultsindicatethatthiskitcanbeusedtodeterminerelativemassvaluesfornaturalhumanVEGF.2Forresearchuseonly.Notforuseindiagnosticprocedures.PRINCIPLEOFTHEASSAYThisassayemploysthequantitativesandwichenzymeimmunoassaytechnique.AmonoclonalantibodyspecificforVEGFhasbeenpre-coatedontoamicroplate.StandardsandsamplesarepipettedintothewellsandanyVEGFpresentisboundbytheimmobilizedantibody.Afterwashingawayanyunboundsubstances,anenzyme-linkedpolyclonalantibodyspecificforVEGFisaddedtothewells.Followingawashtoremoveanyunboundantibody-enzymereagent,asubstratesolutionisaddedtothewellsandcolordevelopsinproportiontotheamountofVEGFboundintheinitialstep.Thecolordevelopmentisstoppedandtheintensityofthecolorismeasured.LIMITATIONSOFTHEPROCEDUREFORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.Thekit首ldnotbeusedbeyondtheexpirationdateonthekitlabel.Donotmixorsubstitutereagentswiththosefromotherlotsorsources.ItisimportantthattheCalibratorDiluentselectedforthestandardcurvebeconsistentwiththesamplesbeingassayed.Ifsamplesgeneratevalueshigherthanthehigheststandard,dilutethesampleswiththeappropriateCalibratorDiluentandrepeattheassay.Anyvariationinstandarddiluent,operator,pipettingtechnique,washingtechnique,incubationtimeortemperature,andkitagecancausevariationinbinding.Variationsinsamplecollection,processing,andstoragemaycausesamplevaluedifferences.Thisassayisdesignedtoeliminateinterferencebysolublereceptors,bindingproteins,andotherfactorspresentinbiologicalsamples.UntilallfactorshavebeentestedintheQuantikineImmunoassay,thepossibilityofinterferencecannotbeexcluded.TECHNICALHINTSWhenmixingorreconstitutingproteinsolutions,alwaysavoidfoaming.Toavoidcross-contamination,changepipettetipsbetweenadditionsofeachstandardlevel,betweensampleadditions,andbetweenreagentadditions.Also,useseparatereservoirsforeachreagent.Toensureaccurateresults,properadhesionofplatesealersduringincubationstepsisnecessary.Whenusinganautomatedplatewasher,addinga30secondsoakperiodfollowingtheadditionofWashBuffer,and/orrotatingtheplate180degreesbetweenwashstepsmayimproveassayprecision.SubstrateSolution首ldremaincolorlessuntiladdedtotheplate.KeepSubstrateSolutionprotectedfromlight.SubstrateSolution首ldchangefromcolorlesstogradationsofblue.StopSolution首ldbeaddedtotheplateinthesameorderastheSubstrateSolution.ThecolordevelopedinthewellswillturnfrombluetoyellowuponadditionoftheStopSolution.WellsthataregreenincolorindicatethattheStopSolutionhasnotmixedthoroughlywiththeSubstrateSolution.www.RnDSystems.com3MATERIALSPROVIDED&STORAGECONDITIONSStoretheunopenedkitat2-8°C.Donotusepastkitexpirationdate.PARTPART#CATALOG#DVE00CATALOG#SVE00DESCRIPTIONSTORAGEOFOPENED/RECONSTITUTEDMATERIALVEGFMicroplate8902181plate6plates96wellpolystyrenemicroplate（12stripsof8wells）coatedwithamousemonoclonalantibodyagainstVEGF.Returnunusedwellstothefoilpouchcontainingthedesiccantpack.Resealalongentireedgeofzip-seal.Maybestoredforupto1monthat2-8°C.*VEGFStandard8902203vials18vials2000pg/vialofrecombinantVEGF165inabufferedproteinbasewithpreservatives;lyophilized.DiscardtheVEGFstocksolutionanddilutionsafter4hours.Useafreshstandardforeachassay.VEGFConjugate8902191vial6vials21mL/vialofapolyclonalantibodyagainstVEGFconjugatedtohorseradishperoxidasewithpreservatives.Maybestoredforupto1monthat2-8°C.*AssayDiluentRD1W8951171vial6vials11mL/vialofabufferedproteinbasewithpreservatives.CalibratorDiluentRD5K8951191vial6vials21mL/vialofabufferedproteinbasewithpreservatives.Forcellculturesupernatesamples.CalibratorDiluentRD6U8951481vial6vials21mL/vialofanimalserumwithpreservatives.Forserum/plasmasamples.WashBufferConcentrate8950031vial6vials21mL/vialofa25-foldconcentratedsolutionofbufferedsurfactantwithpreservatives.ColorReagentA8950001vial6vials12mL/vialofstabilizedhydrogenperoxide.ColorReagentB8950011vial6vials12mL/vialofstabilizedchromogen（tetramethylbenzidine）.StopSolution8950321vial6vials6mL/vialof2Nsulfuricacid.PlateSealersN/A4strips24stripsAdhesivestrips.*Providedthisiswithintheexpirationdateofthekit.DVE00containssufficientmaterialstorunanELISAonone96wellplate.SVE00（SixPak）containssufficientmaterialstorunELISAsonsix96wellplates.ThiskitisalsoavailableinaPharmPak（R&DSystems,Catalog#PDVE00）.PharmPakscontainsufficientmaterialstorunELISAson50microplates.Specificvialcountsofeachcomponentmayvary.Pleaserefertotheliteratureaccompanyingyourorderforspecificvialcounts.4Forresearchuseonly.Notforuseindiagnosticprocedures.OTHERSUPPLIESREQUIREDMicroplatereadercapableofmeasuringabsorbanceat450nm,withthecorrectionwavelengthsetat540nmor570nm.Pipettesandpipettetips.Deionizedordistilledwater.Squirtbottle,manifolddispenser,orautomatedmicroplatewasher.500mLgraduatedcylinder.Polypropylenetesttubesfordilutionofstandards.HumanVEGFControls（optional;availablefromR&DSystems）.PRECAUTIONSCalibratorDiluentRD6Ucontainssodiumazidewhichmayreactwithleadandcopperplumbingtoformexplosivemetallicazides.Flushwithlargevolumesofwaterduringdisposal.VEGFisdetectableinsaliva.Takeprecautionarymeasurestopreventcontaminationofthekitreagentswhilerunningtheassay.TheStopSolutionprovidedwiththiskitisanacidsolution.Wearprotectivegloves,clothing,eye,andfaceprotection.Washhandsthoroughlyafterhand領(lǐng).SAMPLECOLLECTION&STORAGEThesamplecollectionandstorageconditionslistedbelowareintendedasgeneralguidelines.Samplestabilityhasnotbeenevaluated.CellCultureSupernates-Cellculturesupernates首ldcontainatleast1%fetalcalfserumforstabilityoftheVEGF.Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat≤-20°C.Avoidrepeatedfreeze-thawcycles.Serum-Useaserumseparatortube（SST）andallowsamplestoclotfor30minutesbeforecentrifugationfor15minutesat1000xg.Removeserumandassayimmediatelyoraliquotandstoresamplesat≤-20°C.Avoidrepeatedfreeze-thawcycles.Plasma-CollectplasmausingEDTA,heparin,orcitrateasananticoagulant.Centrifugefor15minutesat1000xgwithin30minutesofcollection.Assayimmediatelyoraliquotandstoresamplesat≤-20°C.Avoidrepeatedfreeze-thawcycles.www.RnDSystems.com5REAGENTPREPARATIONBringallreagentstoroomtemperaturebeforeuse.WashBuffer-Ifcrystalshaveformedintheconcentrate,warmtoroomtemperatureandmixgentlyuntilthecrystalshavecompletelydissolved.Dilute20mLofWashBufferConcentrateintodeionizedordistilledwatertoprepare500mLofWashBuffer.SubstrateSolution-ColorReagentsAandB首ldbemixedtogetherinequalvolumeswithin15minutesofuse.Protectfromlight.200μLoftheresultantmixtureisrequiredperwell.VEGFStandard-ReconstitutetheVEGFStandardwith1.0mLofCalibratorDiluentRD5K（forcellculturesupernatesamples）orCalibratorDiluentRD6U（forserum/plasmasamples）.Thisreconstitutionproducesastocksolutionof2000pg/mL.Allowthestandardtositforaminimumof15minuteswithgentleagitationpriortomakingdilutions.ForCellCultureSupernateSamples:Usepolypropylenetubes.Pipette500μLofCalibratorDiluentRD5Kintoeachtube.Usethestocksolutiontoproduceadilutionseries（below）.Mixeachtubethoroughlybeforethenexttransfer.The1000pg/mLdilutionservesasthehighstandard.CalibratorDiluentRD5Kservesasthezerostandard（0pg/mL）.500μLStd.2000pg/mL1000pg/mL500pg/mL250pg/mL125pg/mL62.5pg/mL31.2pg/mL15.6pg/mL500μL500μL500μL500μL500μL500μL500μLStd.2000pg/mL1000pg/mL500pg/mL250pg/mL125pg/mL62.5pg/mL31.2pg/mL500μL500μL500μL500μL500μLForSerum/PlasmaSamples:Usepolypropylenetubes.Pipette500μLofCalibratorDiluentRD6Uintoeachtube.Usethestocksolutiontoproduceadilutionseries（below）.Mixeachtubethoroughlybeforethenexttransfer.Theundilutedstandardservesasthehighstandard（2000pg/mL）.CalibratorDiluentRD6Uservesasthezerostandard（0pg/mL）6Forresearchuseonly.Notforuseindiagnosticprocedures.ASSAYPROCEDUREBringallreagentsandsamplestoroomtemperaturebeforeuse.Itisrecommendedthatallstandards,samples,andcontrolsbeassayedinduplicate.1.Prepareallreagents,workingstandards,andsamplesasdirectedintheprevioussections.2.Removeexcessmicroplatestripsfromtheplateframe,returnthemtothefoilpouchcontainingthedesiccantpack,andreseal.3.ForCellCultureSupernateSamples:Add50μLofAssayDiluentRD1Wtoeachwell.ForSerum/PlasmaSamples:Add100μLofAssayDiluentRD1Wtoeachwell.4.ForCellCultureSupernateSamples:Add200μLofStandard,control,orsampleperwell.ForSerum/PlasmaSamples:Add100μLofStandard,control,orsampleperwell.Coverwiththeadhesivestripprovidedandincubatefor2hoursatroomtemperature.Aplatelayoutisprovidedtorecordthestandardsandsamplesassayed.5.Aspirateeachwellandwash,repeatingtheprocesstwiceforatotalofthreewashes.Washbyfil領(lǐng)eachwellwithWashBuffer（400μL）usingasquirtbottle,manifolddispenser,orautowasher.Completeremovalofliquidateachstepisessentialtogoodperformance.Afterthelastwash,removeanyremainingWashBufferbyaspiratingordecanting.Inverttheplateandblotitagainstcleanpapertowels.6.Add200μLofVEGFConjugatetoeachwell.Coverwithanewadhesivestrip.Incubatefor2hoursatroomtemperature.7.Repeattheaspiration/washasinstep5.8.Add200μLofSubstrateSolutiontoeachwell.Protectfromlight.ForCellCultureSupernateSamples:Incubatefor20minutesatroomtemperature.ForSerum/PlasmaSamples:Incubatefor25minutesatroomtemperature.9.Add50μLofStopSolutiontoeachwell.Ifcolorchangedoesnotappearuniform,gentlytaptheplatetoensurethoroughmixing.Ifthecolorinthewellsisgreenorthecolorchangedoesnotappearuniform,gentlytaptheplatetoensurethoroughmixing.10.Determinetheopticaldensityofeachwellwithin30minutes,usingamicroplatereadersetto450nm.Ifwavelengthcorrectionisavailable,setto540nmor570nm.Ifwavelengthcorrectionisnotavailable,subtractreadingsat540nmor570nmfromthereadingsat450nm.Thissubtractionwillcorrectforopticalimperfectionsintheplate.Readingsmadedirectlyat450nmwithoutcorrectionmaybehigherandlessaccurate.www.RnDSystems.com7CALCULATIONOFRESULTSAveragetheduplicatereadingsforeachstandard,control,andsampleandsubtracttheaveragezerostandardopticaldensity.Createastandardcurvebyreducingthedatausingcomputersoftwarecapableofgeneratingafourparameterlogistic（4-PL）curvefit.Asanalternative,constructastandardcurvebyplottingthemeanabsorbanceforeachstandardonthey-axisagainsttheconcentrationonthex-axisanddrawabestfitcurvethroughthepointsonthegraph.ThedatamaybelinearizedbyplottingthelogoftheVEGFconcentrationsversusthelogoftheO.D.andthebestfitlinecanbedeterminedbyregressionanalysis.Thisprocedurewillproduceanadequatebutlessprecisefitofthedata.Ifsampleshavebeendiluted,theconcentrationreadfromthestandardcurvemustbemultipliedbythedilutionfactor.TYPICALDATAThesestandardcurvesareprovidedfordemonstrationonly.Astandardcurve首ldbegeneratedforeachsetofsamplesassayed.（pg/mL）O.D.AverageCorrected00.0740.0750.07615.60.1180.1200.0450.12131.20.1590.1590.0840.15962.50.2460.2440.1690.2421250.3840.3810.3060.3782500.6660.6680.5930.6695001.2581.2601.1851.26310002.3022.2682.1932.233（pg/mL）O.D.AverageCorrected00.0680.0700.07131.20.1070.1080.0380.11062.50.1490.1510.0810.1531250.2300.2300.1600.2302500.3770.3820.3120.3875000.6570.6780.6080.69910001.2611.2711.2011.28120002.1592.2022.1322.246CALIBRATORDILUENTRD5KCALIBRATORDILUENTRD6U8Forresearchuseonly.Notforuseindiagnosticprocedures.PRECISIONIntra-assayPrecision（Precisionwithinanassay）Threesamplesofknownconcentrationweretestedtwentytimesononeplatetoassessintraassayprecision.Inter-assayPrecision（Precisionbetweenassays）Threesamplesofknownconcentrationweretestedinfortyseparateassaystoassessinterassayprecision.CELLCULTURESUPERNATEASSAYIntra-AssayPrecisionInter-AssayPrecisionSample123123n202020404040Mean（pg/mL）29.112353132.8128495Standarddeviation1.95.018.42.86.433.0CV（%）6.54.13.58.55.06.7SERUM/PLASMAASSAYIntra-AssayPrecisionInter-AssayPrecisionSample123123n202020404040Mean（pg/mL）53.723591064.52501003Standarddeviation3.610.646.25.717.461.7CV（%）6.74.55.18.87.06.2RECOVERYTherecoveryofVEGFspikedtothreedifferentlevelsthroughouttherangeoftheassayinvariousmatriceswasevaluated.SampleTypeAverage%RecoveryRangeCellculturemedia（n=5）10295-111%Serum（n=5）10292-115%EDTAplasma（n=5）9782-113%Heparinplasma（n=5）9382-102%Citrateplasma（n=5）10088-113%SENSITIVITYUsingCalibratorDiluentRD5Ktheminimumdetectabledose（MDD）ofVEGFistypicallylessthan5.0pg/mL.UsingCalibratorDiluentRD6UtheMDDistypicallylessthan9.0pg/mL.TheMDDwasdeterminedbyaddingtwostandarddeviationstothemeanopticaldensityvalueoftwentyzerostandardreplicatesandcalculatingthecorrespondingconcentration.www.RnDSystems.com9LINEARITYToassesslinearityoftheassay,sampleswerespikedwithhighconcentrationsofVEGFanddilutedwiththeappropriateCalibratorDiluenttoproducesampleswithvalueswithinthedynamicrangeoftheassay.Cellculturemedia（n=5）Serum（n=5）EDTAplasma（n=5）Heparinplasma（n=5）Citrateplasma（n=5）1:2Average%ofExpected9897979495Range（%）94-10091-10382-10787-9990-1001:4Average%ofExpected9697989394Range（%）93-9993-10491-10685-9889-991:8Average%ofExpected9396969292Range（%）88-10293-10389-10685-10185-971:16Average%ofExpected9394949492Range（%）88-10591-10184-10683-10385-98CALIBRATIONThisimmunoassayiscalibratedagainstahighlypurifiedSf21-expressedrecombinanthumanVEGF165producedatR&DSystems.TheNIBSC/WHOVEGF165preparation02/286（recombinanthumanDNA）wasevaluatedinthiskit.Thedoseresponsecurveofthestandard02/286parallelstheQuantikinestandardcurve.ToconvertsamplevaluesobtainedwiththeQuantikineHumanVEGFkittoapproximateNIBSC/WHO02/286Units,usetheequationbelow.NIBSC/WHO（02/286）approximatevalue（U/mL）=0.002xQuantikineVEGFvalue（pg/mL）Note:BasedondatageneratedinApril2011.10Forresearchuseonly.Notforuseindiagnosticprocedures.SAMPLEVALUESSerum/Plasma-SamplesfromapparentlyhealthyvolunteerswereevaluatedforthepresenceofVEGFinthisassay.Nomedicalhistorieswereavailableforthedonorsusedinthisstudy.SampleTypeMeanofDetectable（pg/mL）%DetectableRange（pg/mL）Serum（n=37）22010062-707EDTAplasma（n=37）6124ND-115Heparinplasma（n=37）4122ND-55Citrateplasma（n=37）___0NDND=Non-detectableCellCultureSupernates-Humanperipheralbloodmononuclearcells（1x106cells/mL）wereculturedinRPMIsupplementedwith5%fetalcalfserum,50μMβ-mercaptoethanol,2mML-glutamine,100U/mLpenicillin,and100μg/mLstreptomycinsulfate.Thecellswereculturedunstimulatedorstimulatedwith10μg/mLPHAfor1and5days.AliquotsofthecellculturesupernateswereremovedandassayedforlevelsofnaturalVEGF.ConditionDay1（pg/mL）Day5（pg/mL）Unstimulated356332Stimulated141440www.RnDSystems.com11SPECIFICITYThisassayrecognizesnaturalandrecombinanthumanVEGF.ThisassayalsorecognizesrecombinanthumanVEGF165b.Thefactorslistedbelowwerepreparedat50ng/mLinCalibratorDiluentandassayedforcrossreactivity.Preparationsofthefollowingfactorsat50ng/mLinamid-rangeVEGFcontrolwereassayedforinterference.Thefollowingfactorsshowednocross-reactivityorinterference.Recombinanthuman:PDGF-AAPDGF-ABPDGF-BBPDGF-CCPDGF-DDPlGFPlGF-2VEGF165/PlGFVEGF-B167VEGF-CVEGF-DVEGFR3Recombinantmouse:PDGF-CCPlGF-2VEGF120VEGF164VEGFR3Recombinantrat:PDGF-AAPDGF-ABPDGF-BBVEGF164Recombinantzebrafish:VEGFNaturalproteins:humanPDGFporcinePDGFVEGF-relatedfactorsshowingcross-reactivityorinterference.RecombinanthumanVEGFR1/Flt-1Interferenceatlevels≥500pg/mLRecombinanthumanVEGFR2/KDRInterferenceatlevels≥2000pg/mLRecombinantmouseVEGFR1/Flk-1Interferenceatlevels≥500pg/mLRecombinantmouseVEGFR2/KDRInterferenceatlevels≥4000pg/mLRecombinantcanineVEGFCross-reactsapproximately67%RecombinantfelineVEGFCross-reactsapproximately82%12Forresearchuseonly.Notforuseindiagnosticprocedures.REFERENCES1.Leung,D.W.etal.（1989）Science246:1306.2.Keck,P.J.etal.（1989）Science246:1309.3.Byrne,A.M.etal.（2005）J.Cell.Mol.Med.9:777.4.Robinson,C.J.andS.E.Stringer（2001）J.Cell.Sci.114:853.5.Richardson,R.S.etal.（1999）Am.J.Physiol.277:H2247.6.Sugishita,Y.etal.（2000）Biochem.Biophys.Res.Commun.268:657.7.Yamane,A.etal.（1994）Oncogene9:2683.8.Goad,D.L.etal.（1996）Endocrinology137:2262.9.Gaudry,M.etal.（1997）Blood90:4153.10.Mclaren,J.etal.（1996）J.Clin.Invest.98:482.11.Diaz,B.V.etal.（2000）J.Biol.Chem.275:642.12.Asano,A.etal.（1997）Biochem.J.328:179.13.Bautz,F.etal.（2000）Exp.Hematol.28:700.14.Namiki,A.etal.（1995）J.Biol.Chem.270:31189.15.Nauck,M.etal.（1997）Am.J.Respir.Cell.Mol.Biol.16:398.16.Angelo,L.S.andR.Kurzrock（2007）Clin.CancerRes.13:2825.17.Neufeld,G.etal.（1999）FASEB.J.13:9.18.Kowalewski,M.P.etal.（2005）Accession#ABB82619.19.Pan,Q.etal.（2007）J.Biol.Chem.282:24049.20.Dai,J.andA.B.Rabie（2007）J.Dent.Res.86:937.21.Breier,G.（2000）Semin.Thromb.Hemost.26:553.22.Barleon,B.etal.（1996）Blood87:3336.23.Weis,S.M.andD.A.Cheresh（2005）Nature437:497.24.Thurston,G.（2002）J.Anat.200:575.25.Grothey,A.andE.Galanis（2009）Nat.Rev.Clin.Oncol.6:507.26.Carvalho,J.F.etal.（2007）J.Clin.Immunol.27:246.www.RnDSystems.com13PLATELAYOUTUsethisplatelayouttorecordstandardsandsamplesassayed.14Forresearchuseonly.Notforuseindiagnosticprocedures.[詳細]
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- 人原鈣黏素1（PCDH1）ELISAKit人白介素2受體（IL-2R）ELISAKit人皮膚T細胞虜獲趨化因子（CTACK/CCL27）ELISAKit人胸腎表達趨化因子（BRAK/CXCL14）ELISAKit人B-淋巴細胞趨化因子1（BLC-1/CXCL13）ELISAKit人結(jié)締組織活化肽Ⅲ（CTAPⅢ）ELISAKit人巨噬細胞集落刺激因子受體（M-CSFR）ELISAKit人免疫球蛋白AFc段受體Ⅰ（FcαRⅠ/CD89）ELISAKit人免疫球蛋白EFc段受體Ⅱ（FcεRⅡ/CD23）ELISAKit人免疫球蛋白GFc段受體Ⅲ（FcγRⅢ/CD16）ELISAKit人免疫球蛋白GFc段受體Ⅱ（FcγRⅡ/CD32）ELISAKit人免疫球蛋白GFc段受體Ⅰ（FcγRⅠ/CD64）ELISAKitGFc段受體Ⅰ（FcγRⅠ/CD64）ELISAKit人免疫球蛋白GFc段受體Ⅲ（FcγRⅢ/CD16）ELISAKit人免疫球蛋白GFc段受體Ⅱ（FcγRⅡ/CD32）ELISAKit人免疫球蛋白GFc段受體Ⅰ（FcγRⅠ/CD64）ELISAKit人粒細胞趨化蛋白-2（GCP-2/CXCL6）ELISAKit人ω干擾素（IFN-ω）ELISAKit人糖基化依賴的細胞黏附分子（GlyCAM-1）ELISAKit人干擾素調(diào)節(jié)因子（IRF）ELISAKit人淋巴毒素β（LTB）ELISAKit人淋巴毒素α（LTA）ELISAKit人CC趨化因子受體1（CCR1）ELISAKit人CX3C趨化因子受體1（CX3CR1）ELISAKit人肺部活化調(diào)節(jié)趨化因子（PARC/CCL18）ELISAKit人黏膜地址素細胞黏附分子（MAdCAM-1）ELISAKit人β干擾素（IFN-β/IFNB）ELISAKit人可溶性CD38（sCD38）ELISAKit人可溶性CD21（CR2/sCD21）ELISAKit人可溶性瘦素受體（sLR）ELISAKit人Toll樣受體9（TLR-9/CD289）ELISAkit人轉(zhuǎn)化生長因子β2（TGFβ2）ELISAKit人單核細胞趨化蛋白4（MCP-4/CCL13）ELISAKit人白三烯D4（LTD4）ELISAKit人N鈣黏蛋白/神經(jīng)鈣黏蛋白（N-Cad）ELISAKit人肝素結(jié)合性表皮生長因子（HB-EGF）ELISAKit人紅細胞刺激因子（ESF）ELISAKit人腫瘤壞死因子相關(guān)激活誘導(dǎo)因子（TRANCE）ELISAKit人生長激素釋放因子（GH-RF）ELISAKit人巨噬細胞趨化因子（MCF）ELISAKit人α/β干擾素受體（IFN-α/βR）ELISAKit人B細胞生長因子（BCGF）ELISAKit人B細胞分化因子（BCDF）ELISAKit人上皮細胞粘附分子（Ep-CAM/CD362）ELISAKit人可溶性粘附分子（Sam）ELISAKit人巨噬細胞替代激活相關(guān)化學因子1（AmAC-1）ELISAKit人可溶性血管內(nèi)皮生長因子受體2（VEGFR-2/sFLK-1）ELISAKit人胸腺基質(zhì)淋巴細胞生成素（TSLP）ELISAKit人穿孔素/成孔蛋白（PF/PFP）ELISAKit人多效生長因子（PTN）ELISAKit人可溶性CD28（sCD28）ELISAKit人淋巴細胞因子ELISAKit人胸腺活化調(diào)節(jié)趨化因子（TARC/CCL17）ELISAKit人神經(jīng)細胞粘附分子配體1（NCAM-L1/CD171）ELISAKit人神經(jīng)保護因子（CVNPF）ELISAKit人可溶性腫瘤壞死因子α受體（sTNFαR）ELISAKit人可溶性細胞因子受體（sCKR）ELISAKit人可溶性凋亡相關(guān)因子配體（sFASL）ELISAKit人細胞凋亡YZ因子（IAP）ELISAKit人集落刺激因子（CSF）ELISAKit人γ干擾素誘導(dǎo)單核細胞因子（MIGF/CXCL9）ELISAKit人干擾素誘導(dǎo)T細胞趨化因子（ITAC/CXCL11）ELISAKit人CD14分子（CDl4）ELISAKit人凋亡誘導(dǎo)因子（AIF）ELISAKit人白細胞共同抗原（LCA/CD45）ELISAKit人CD4分子（CD4）ELISAKit人P鈣黏蛋白/胎盤鈣黏蛋白（P-cad）ELISAKit人角化細胞生長因子（KGF）ELISAKit人血小板衍生生長因子BB（PDGF-BB）ELISAKit人CXC趨化因子配體16（CXCL16）ELISAKit人CXC趨化因子受體3（CXCR3）ELISAKit人γ干擾素誘導(dǎo)蛋白16/p16（IFI16/p16）ELISAKit人基質(zhì)細胞衍生因子1a（SDF-1a/CXCL12）ELISAKit人淋巴細胞趨化因子（Lptn/LTN/XCL1）ELISAKit人α干擾素（IFN-α）ELISAKit人可溶性CD86（B7-2/sCD86）ELISAKit人白介素27（IL-27）ELISAKit人白介素23（IL-23）ELISAKit人巨噬細胞移動YZ因子（MIF）ELISAKit人組織因子途徑Y(jié)Z物（TFPI）ELISAKit人干擾素誘導(dǎo)蛋白10（IP-10/CXCL10）ELISAKit人白介素1（IL-1）ELISAKit人白介素17（IL-17）ELISAKit人白介素1β（IL-1β）ELISAKit人表皮生長因子（EGF）ELISAKit人堿性成纖維細胞生長因子（bFGF）ELISAKit人巨噬細胞炎性蛋白5（MIP-5）ELISAKit人可溶性E選擇素（sE-selectin）ELISAKit人可溶性細胞間粘附分子1（sICAM-1）ELISAKit人細胞間粘附分子2（ICAM-2/CD102）ELISAKit人細胞間粘附分子3（ICAM-3/CD50）ELISAKit人結(jié)締組織生長因子（CTGF）ELISAKit人白介素18（IL-18）ELISAKit人粘膜相關(guān)上皮趨化因子（MEC/CCL28）ELISAKit人粘膜相關(guān)上皮趨化因子（MEC/CCL28）ELISAKit人B細胞活化因子受體（BAFF-R）ELISAKit人血管內(nèi)皮細胞生長因子受體3（VEGFR-3/Flt-4）ELISAKit人血管內(nèi)皮細胞生長因子受體1（VEGFR-1/Flt1）ELISAKit人血管內(nèi)皮細胞生長因子D（VEGF-D）ELISAKit[詳細]
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2024-09-30 09:18
產(chǎn)品樣冊
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- Monkey ELISA Kit
- MonkeyELISAKit[詳細]
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2024-09-28 07:04
產(chǎn)品樣冊
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- 上海易利為您提供Human GST elisa kit 英文說明書
- 本公司為ELISA試劑盒供應(yīng)商，價格公正，售前，售中，售后全程為您服務(wù)。提供免費代測服務(wù)，一周內(nèi)出結(jié)果，保證質(zhì)量歡迎來電咨詢!許經(jīng)理021-6051540918217407991[詳細]
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2018-10-01 10:00
產(chǎn)品樣冊
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- 人凝血酶ELISA Kit
- 人凝血酶ELISA Kit[詳細]
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2014-08-21 00:00
安裝說明
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- Mouse CaSR ELISA kit
- Mouse CaSR ELISA kit[詳細]
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2015-10-09 00:00
操作手冊
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- Rat Aβ1-42 ELISA kit
- www.biokanu.comRatamyloidbetapeptide1-42（Aβ1-42）ELISAKitProd.No.3R285FROM:RBForthequantitativeinvitrodeterminationofAβ1-42concentrationsinRatsupernates,serum,plasmaandtissue.FORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.TABLEOFCONTENTSContentsPageTABLEOFCONTENTS..2INTENDEDUSE..3PRINCIPLE..3WARNINGSANDPRECAUTIONS..4MATERIALSPROVIDEDWITHTHEKIT.7MATERIALSREQUIREDBUTNOTPROVIDED..7STORAGECONDITIONS..8REAGENTPREPARATION..9SPECIMENCOLLECTIONANDPREPARATION..9ASSAYPROCEDURE..10CALCULATIONOFRESULTS..13REFERENCES..14INTENDEDUSEAnenzymeimmunoassayforthequantitativeinvitrodiagnosticmeasurementofRatAβ1-42incellculturesupernates,serum,plasmaandtissue.PRINCIPLEThekitassayRatAβ1-42levelinthesample，usePurifiedRatAβ1-42antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddAβ1-42towells,CombinedAβ1-42antibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofAβ1-42inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.WARNINGSANDPRECAUTIONSlThiskitisforinvitrodiagnosticuseonly.Forprofessionaluseonly.lAllreagentsofthistestkitwhichcontainhumanserumorplasmahavebeentestedandconfirmednegativeforHIVI/II,HBsAgandHCVbyFDAapprovedprocedures.Allreagents,however,首ldbetreatedaspotentialbiohazardsinuseandfordisposal.lBeforestartingtheassay,readtheinstructionscompletelyandcarefully.Usethevalidversionofthepackageinsertprovidedwiththekit.Besurethateverythingisunderstood.lThemicroplatecontainssnap-offstrips.Unusedwellsmustbestoredat2°Cto8°Cinthesealedfoilpouchandusedintheframeprovided.lPipettingofsamplesandreagentsmustbedoneasquicklyaspossibleandinthesamesequenceforeachstep.lUsereservoirsonlyforsinglereagents.Thisespeciallyappliestothesubstratereservoirs.Usingareservoirfordispensingasubstratesolutionthathadpreviouslybeenusedfortheconjugatesolutionmayturnsolutioncolored.Donotpourreagentsbackintovialsasreagentcontaminationmayoccur.lMixthecontentsofthemicroplatewellsthoroughlytoensuregoodtestresults.Donotreusemicrowells.lDonotletwellsdryduringassay;addreagentsimmediatelyaftercompletingtherinsingsteps.lAllowthereagentstoreachroomtemperature（21-26°C）beforestartingthetest.Temperaturewillaffecttheabsorbancereadingsoftheassay.However,valuesforthepatientsampleswillnotbeaffected.lNeverpipetbymouthandavoidcontactofreagentsandspecimenswithskinandmucousmembranes.lDonotsmoke,eat,drinkorapplycosmeticsinareaswherespecimensorkitreagentsarehandled.lWeardisposablelatexgloveswhenhand領(lǐng)specimensandreagents.Microbialcontaminationofreagentsorspecimensmaygivefalseresults.lHand領(lǐng)首ldbedoneinaccordancewiththeproceduresdefinedbyanappropriatenationalbiohazardsafetyguidelineorregulation.lDonotusereagentsbeyondexpirydateasshownonthekitlabels.lAllindicatedvolumeshavetobeperformedaccordingtotheprotocol.Optimaltestresultsareonlyobtainedwhenusingcalibratedpipettesandmicrotiterplatereaders.lDonotmixorusecomponentsfromkitswithdifferentlotnumbers.Itisadvisednottoexchangewellsofdifferentplatesevenofthesamelot.Thekitsmayhavebeenshippedorstoredunderdifferentconditionsandthebindingcharacteristicsoftheplatesmayresultslightlydifferent.lAvoidcontactwithStopSolutioncontaining0.5MH2SO4.Itmaycauseskinirritationandburns.lSomereagentscontainProclin,BNDand/orMITaspreservatives.Incaseofcontactwitheyesorskin,flushimmediatelywithwater.lTMBsubstratehasanirritanteffectonskinandmucosa.Incaseofpossiblecontact,washeyeswithanabundantvolumeofwaterandskinwithsoapandabundantwater.Washcontaminatedobjectsbeforereusingthem.Ifinhaled,takethepersontoopenair.lChemicalsandpreparedorusedreagentshavetobetreatedashazardouswasteaccordingtothenationalbiohazardsafetyguidelineorregulation.lForinformationonhazardoussubstancesincludedinthekitpleaserefertoMaterialSafetyDataSheetsMATERIALSPROVIDEDWITHTHEKITMaterialsprovidedwiththekit96determinationsStorageUsermanual1Closureplatemembrane2Sealedbags1Microelisastripplate12-8℃Standard：900pg/ml0.5ml×1bottle2-8℃Standarddiluent1.5ml×1bottle2-8℃HRP-Conjugatereagent6ml×1bottle2-8℃Samplediluent6ml×1bottle2-8℃ChromogenSolutionA6ml×1bottle2-8℃ChromogenSolutionB6ml×1bottle2-8℃StopSolution6ml×1bottle2-8℃washsolution30×20ml×1bottle2-8℃MATERIALSREQUIREDBUTNOTPROVIDEDlMicroplatereadercapableofmeasuringabsorbanceat450nm.lPrecisionpipettestodeliver2mlto1mlvolumes.l100mland1litergraduatedcylinders.lCalibratedadjustableprecisionpipettes,preferablywithdisposableplastictips.（Amanifoldmulti-channelpipetteisdesirableforlargeassays.）lAbsorbentpaper.l37°Cincubator.lDistilledordeionizedwater.lDataanalysisandgraphingsoftware.Graphpaper:linear（Cartesian）,log-logorsemi-log,orlog-logitasdesired.lTubestopreparestandardorsampledilutions.STORAGECONDITIONSuWhenstoredat2°Cto8°Cunopenedreagentswillretainreactivityuntilexpirationdate.uDonotusereagentsbeyondthisdate.Openedreagentsmustbestoredat2°Cto8°C.uMicrotiterwellsmustbestoredat2°Cto8°C.Oncethefoilbaghasbeenopened,care首ldbetakentocloseittightlyagain.uOpenedkitsretainactivityfor8weeksifstoredasdescribedabove.REAGENTPREPARATIONBringallreagentstoroomtemperaturebeforeuseSPECIMENCOLLECTIONANDPREPARATIONSerum-Useaserumseparatortube（SST）andallowsamplestoclotfor30minutesbeforecentrifugationfor15minutesatapproximately1000xg.Removeserumandassayimmediatelyoraliquotandstoresamplesat-20°Cor-80°C.Plasma-CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor15minutesat1000xgat2-8°Cwithin30minutesofcollection.Storesamplesat-20°Cor-80°C.Avoidrepeatedfreeze-thawcycles.Cellculturefluidandotherbiologicalfluids-Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat-20°Cor-80°C.Avoidrepeatedfreeze-thawcyclesASSAYPROCEDUREuGeneralRemarkslAllreagentsandspecimensmustbeallowedtocometoroomtemperaturebeforeuse.Allreagentsmustbemixedwithoutfoaming.lOncethetesthasbeenstarted,allsteps首ldbecompletedwithoutinterruption.lUsenewdisposalplasticpipettetipsforeachstandard,controlorsampleinordertoavoidcrosscontamination.lAbsorbanceisafunctionoftheincubationtimeandtemperature.Beforestartingtheassay,itisrecommendedthatallreagentsareready,capsremoved,allneededwellssecuredinholder,etc.Thiswillensureequalelapsedtimeforeachpipettingstepwithoutinterruption.lAsageneralruletheenzymaticreactionislinearlyproportionaltotimeandtemperature.lDetermineabsorptionwithanELISAreaderat450nmagainst620nmasreference.Ifnoreferencewavelengthisavailable,readonlyat450nm.Iftheextinctionofthehigheststandardexceedsthemeasurementrangeofthephotometer,absorptionmustbemeasuredimmediatelyat405nmagainst620nmasreference.uAssayProcedure1.DiluteandaddsampletoStandard:set10StandardwellsontheELISAplatescoated,addStandard100μltothefirstandthesecondwell,thenaddStandarddilution50μltothefirstandthesecondwell,mix;takeout100μlformthefirstandthesecondwellthenaddittothethirdandtheforthwellseparately.thenaddStandarddilution50μltothethirdandtheforthwell,mix;thentakeout50μlfromthethirdandtheforthwelldiscard,add50μltothefifthandthesixthwell,thenaddStandarddilution50μltothefifthandthesixthwell,mix;takeout50μlfromthefifthandthesixthwellandaddtotheseventhandtheeighthwell,thenaddStandarddilution50μltotheseventhandtheeighthwell,mix;takeout50μlfromtheseventhandtheeighthwellandaddtotheninthandthetenthwell,addStandarddilution50μltotheninthandthetenthwell,mix,takeout50μlfromtheninthandthetenthwelldiscard（addSample50μltoeachwellafterDiluting,（density:600pg/ml，400pg/ml，200pg/ml,100pg/ml，50pg/ml）.50pg/ml100pg/ml600pg/ml200pg/ml900pg/ml400pg/ml2.addsample：Setblankwellsseparately（blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame）.testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl（samplefinaldilutionis5-fold）,addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-foldwashsolutiondiluted30-foldwithdistilledwaterandreserve.5.washing：UncoverClosureplatemembrane,discardLiquid,drybyswing,addwashingbuffertoeverywell,stillfor30sthendrain,repeat5times,drybypat.6.addenzyme：AddHRP-Conjugatereagent50μltoeachwell,exceptblankwell.7.incubate：Operationwith3.8.washing：Operationwith5.9.color：AddChromogenSolutionA50ulandChromogenSolutionBtoeachwell,evadethelightpreservationfor15minat37℃10.Stopthereaction：AddStopSolution50μltoeachwell,Stopthereaction（thebluecolorchangetoyellowcolor）.11.assay：takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionandwithin15min.CALCULATIONOFRESULTSlCalculatetheaverageabsorbancevaluesforeachsetofstandards,controlsandpatientsamples.lConstructastandardcurvebyplottingthemeanabsorbanceobtainedfromeachstandardagainstits.lconcentrationwithabsorbancevalueonthevertical（Y）axisandconcentrationonthehorizontal（X）axis.lUsingthemeanabsorbancevalueforeachsampledeterminethecorrespondingconcentrationfromthestandardcurve.lAutomatedmethod:TheresultsintheIFUhavebeencalculatedautomaticallyusinga4PL.l（4ParameterLogistics）curvefit.4ParameterLogisticsisthepreferredcalculationmethod.Otherdata.lreductionfunctionsmaygiveslightlydifferentresults.lTheconcentrationofthesamplescanbereaddirectlyfromthisstandardcurve.Sampleswith.lconcentrationshigherthanthatofthehigheststandardhavetobefurtherdiluted.Forthecalculationof.ltheconcentrationsthisdilutionfactorhastobetakenintoaccount.REFERENCESREF:Cat.-No.:/Kat.-Nr.:/No.-Cat.:/Cat.-No.:/N.Cat.:/N.CatLOT:Lot-No.:/Chargen-Bez.:/No.Lot:/Lot-No.:/LoteN.:/Lotton.::No.ofTests:/Kitgre:/Nb.deTests:/No.deDeterm.:/N.deTestes:/Quantitàdeitests::Keepawayfromheatordirectsunlight./VorHitzeunddirekterSonneneinstrahlungschützen./Garderàl’abridelachaleuretdetouteexpositionlumineuse./Manténgasealejadodelcalorolaluzsolardirecta./Manterlongedocalorouluzsolardirecta./Nonesporreairaggisolari.:Readinstructionsbeforeuse./Arbeitsanleitunglesen./Lirelafichetechniqueavantemploi./Lealasinstruccionesantesdeusar./Lerasinstruesantesdeusar./Leggereleistruzioniprimadell’uso.:Storeat:/Lagernbei:/Stockerà:/Almacenea:/Armazenara:/Conservarea:[詳細]
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2018-09-22 10:00
產(chǎn)品樣冊
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- Mouse （VLDL）ELISA Kit
- Mouseverylowdensitylipoprotein(VLDL)ELISAKitFORRESEARCHUSEONLY.NotforclinicaldiagnosisuseCATALOG#:DAG859INTRODUCTION?ThiskitallowsforthedeterminationofVLDLconcentrationsinMouseserum?Detectionofspecies:Mouse?Detectionmedium:serum,cellculturesupernates.?Assayrange：6.0μg/ml-160μg/mlPRINCIPLEOFTESTThekitassayMouseVLDLlevelinthesample，usePurifiedMouseVLDLantibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddVLDLtowells,CombinedVLDLantibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofMouseVLDLinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Phone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG6342COMPOSITIONOFTHEKIT1washsolution20ml×1bottle7StopSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard（320μg/ml）0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11Closureplatemembrane26ChromogenSolutionB6ml×1bottle12Sealedbags1STORAGECONDITIONS?Theunopenedkitshallbestoredat[2-8℃].?Foropenedkitcanbestoredat[2-8℃]forupto1month.Ifnotbeusedrecently,thestandard首ldbekeptin-20℃.WASHINGMETHOD?Manuallywashingmethod:shakeawaytheremainedliquidintheenzymeplates;placesomebibulouspapersonthetest-bed,andflaptheplatesontheupsidedownstrongly.Injectatleast0.35mlafter-dilutionwashingsolutionintothewell,andmarinate1~2minutes.Repeatthisprocessaccordingtoyourrequirements.?Automaticwashingmethod:ifthereisautomaticwashingmachine,it首ldonlybeusedinthetestwhenyouarequitefamiliarwithitsfunctionandperformance.SAMPLEPREPARATION1.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.Phone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG63432.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.ASSAYPROCEDUREStep1:Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:Step2:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.Step3:Incubate:Coverwiththeadhesivestripprovided,incubatefor30minat37℃.Step4:Configurateliquid:Dilutewashsolution30-fold(or20-fold)withdistilledwater.Step5:Washing:Uncovertheadhesivestrip,discardliquid,Pipettewashingbuffertoeverywell,stillfor30sthendrain,repeat5times.Step6:Addenzyme:PipetteHRP-Conjugatereagent50μltoeachwell,exceptblankwell.Step7:Incubate:Operationwith3.Step8:Washing:Operationwith5.160μg/ml5Standard150μlOriginaldensityStandard+150μlStandarddiluent80μg/ml4Standard150μl5Standard+150μlStandarddiluent40μg/ml3Standard150μl4Standard+150μlStandarddiluent20μg/ml2Standard150μl3Standard+150μlStandarddiluent10μg/ml1Standard150μl2Standard+150μlStandarddiluentPhone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG6344Step9:Color:PipetteChromogenSolutionA50ulandChromogenSolutionBtoeachwell,avoidthelightpreservationfor15minat37℃Step10:Stopthereaction:PipetteStopSolution50μltoeachwell,Stopthereaction(thebluechangetoyellow).Step11:Calculate:takeblankwellaszero,Readabsorbanceat450nmafterPipetteingStopSolutionwithin15min.CALCULATIONOFRESULTTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,withthesampleODvalueintheequation,calculatethesampledensity,multipliedbythedilutionfactor,theresultisthesampleactualdensity.EXPIRATIONSixmonths[seelabelontheouterboxforthespecificdate].Phone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG634TTENTION?Thekittakesoutfromtherefrigeration首ldbebalanced15-30minutesintheroomtemperature,ifthecoatedELISAplateshavenotbeenusedupafteropening,theplate首ldbestoredinsealedbag.?washingbufferwillCrystallizationseparation,itcanbeheatedthewaterhelpsdissolvewhendilute.Washingdoesnotaffecttheresult.?addSamplewithsamplerEachstep,Andproofreaditsaccuracyfrequently,avoidstheexperimentalerror.addsamplewithin5min,ifthenumberofsampleismuch,recommendtouseVolley.?ifthetestingmaterialcontentisexcessivelyhigher(ThesampleODisbiggerthanthefirststandardwell),pleasediluteSample(n-fold),Pleasediluenteandmultipliedbythedilutionfactor.（×n×5）.?Closureplatemembraneonlylimitsthedisposableuse,toavoidcross-contamination.?Thesubstrate首ldevadethelighttobepreserved.?Pleaserefertotheuserinstructionstrictly,thetestresultdeterminationmusttakethemicrotiterplatereaderasastandard.?Thepreparationofsamplesandallthereagents首ldrefertoinfectivematerialprocess.?Donotmixreagentswiththosefromotherlots[詳細]
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2018-10-31 10:00
產(chǎn)品樣冊
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- Human 8-OHDG elisa試劑盒
- Human 8-OHDG elisa試劑盒[詳細]
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2024-09-16 18:47
期刊論文
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- 人ELISA試劑盒,TPA ELISA Kit
- 96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒相關(guān)產(chǎn)品：96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒IL-10,小鼠白介素-10Elisa試劑盒磺胺二甲基惡唑,標準品JLJL200712人Elisa試劑盒,人ProteinCElisa試劑盒，HumanProteinCELISA試劑盒CAS號:108-69-0,3,5-二,98%人Elisa試劑盒,人CD30Elisa試劑盒，HumanClusterofdifferentiation30,CD30ELISA試劑盒CAS:893-36-7,鹽酸-L-白氨酰-2-萘胺/L-亮氨酰-2-萘胺鹽酸鹽/L-白氨酰-β-萘胺鹽酸鹽/鹽酸-L-亮氨酰-2-萘胺/L（+）-亮氨酰-2-萘基鹽酸氨/L-Leucyl-2-naphthylamidehydrochloride,BR,98%,1克,避光,-20℃96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒Humanbacterialvaginosis,BVELISA試劑盒人（BV）kit說明書,細菌性陰道病Elisa試劑盒CXCR3ELISAKit,大鼠CXC趨化因子受體3Elisa檢測試劑盒蒙花苷,標準品,含量測定,20mg,常溫,避光PorcineapoproteinA1,apo-A1ELISA試劑盒豬（apo-A1）kit說明書,載脂蛋白A1Elisa試劑盒大鼠淋巴細胞因子ELISA試劑盒HumanhepatitisBvirusXinteractingprotein,HBXIPELISAKit人異常凝血酶原（APT）ELISA試劑盒HumanAbnormalprothrombin,APTELISA試劑盒草烏甲素,標準品,含量測定,50mg,常溫,避光96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒GRP1957,營養(yǎng)肉湯,供一般細菌培養(yǎng)、轉(zhuǎn)種和增菌用,250g小鼠生長激素釋放多肽（GHRP）ELISA試劑盒HumanMotilin,MTLELISAKitCAS號:4767-3-7,2,2-雙（羥甲基）丙酸,98%[詳細]
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2018-10-23 10:31
產(chǎn)品樣冊
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- [ELISA]CD8分子（CD8）ELISA Kit
- 羊CD8分子（CD8）試劑盒使用說明書本試劑盒僅供研究使用。檢測范圍：96T7IU/ml-240IU/ml試劑盒組成130倍濃縮洗滌液20ml×1瓶7終止液6ml×1瓶2酶標試劑6ml×1瓶8標準品（400IU/ml）0.5ml×1瓶3酶標包被板12孔×8條9標準品稀釋液1.5ml×1瓶4樣品稀釋液6ml×1瓶10說明書1份5顯色劑A液6ml×1瓶11封板膜2張6顯色劑B液6ml×1/瓶12密封袋1個標本要求1．標本采集后盡早進行提取，提取按相關(guān)文獻進行，提取后應(yīng)盡快進行實驗。若不能馬上進行試驗，可將標本放于-20℃保存，但應(yīng)避免反復(fù)凍融2．不能檢測含NaN3的樣品，因NaN3YZ辣根過氧化物酶的（HRP）活性。羊CD8分子ELISA試劑盒用于測定羊血清、血漿及相關(guān)液體樣本中CD8分子（CD8）含量。操作步驟1.標準品的稀釋：本試劑盒提供原倍標準品一支，用戶可按照下列圖表在小試管中進行稀釋。200IU/ml5號標準品150μl的原倍標準品加入150μl標準品稀釋液100IU/ml4號標準品150μl的5號標準品加入150μl標準品稀釋液50IU/ml3號標準品150μl的4號標準品加入150μl標準品稀釋液25IU/ml2號標準品150μl的3號標準品加入150μl標準品稀釋液12.5IU/ml1號標準品150μl的2號標準品加入150μl標準品稀釋液2.加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標試劑，其余各步操作相同）、標準孔、待測樣品孔。在酶標包被板上標準品準確加樣50μl，待測樣品孔中先加樣品稀釋液40μl，然后再加待測樣品10μl（樣品Z終稀釋度為5倍）。加樣將樣品加于酶標板孔底部，盡量不觸及孔壁，輕輕晃動混勻。3.溫育：用封板膜封板后置37℃溫育30分鐘。4.配液：將30倍濃縮洗滌液用蒸餾水30倍稀釋后備用5.洗滌：小心揭掉封板膜，棄去液體，甩干，每孔加滿洗滌液，靜置30秒后棄去，如此重復(fù)5次，拍干。6.加酶：每孔加入酶標試劑50μl，空白孔除外。7.溫育：操作同3。8.洗滌：操作同5。9.顯色：每孔先加入顯色劑A50μl，再加入顯色劑B50μl，輕輕震蕩混勻，37℃避光顯色15分鐘.10.終止：每孔加終止液50μl，終止反應(yīng)（此時藍色立轉(zhuǎn)黃色）。11.測定：以空白空調(diào)零，450nm波長依序測量各孔的吸光度（OD值）。測定應(yīng)在加終止液后15分鐘以內(nèi)進行。羊CD8分子ELISA試劑盒注意事項1．試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用，酶標包被板開封后如未用完，板條應(yīng)裝入密封袋中保存。2．濃洗滌液可能會有結(jié)晶析出，稀釋時可在水浴中加溫助溶，洗滌時不影響結(jié)果。3．各步加樣均應(yīng)使用加樣器，并經(jīng)常校對其準確性，以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi)，如標本數(shù)量多，推薦使用排槍加樣。4．請每次測定的同時做標準曲線，**做復(fù)孔。如標本中待測物質(zhì)含量過高（樣本OD值大于標準品孔**孔的OD值），請先用樣品稀釋液稀釋一定倍數(shù)（n倍）后再測定，計算時請Z后乘以總稀釋倍數(shù)（×n×5）。5．封板膜只限一次性使用，以避免交叉污染。6．底物請避光保存。7．嚴格按照說明書的操作進行，試驗結(jié)果判定必須以酶標儀讀數(shù)為準.8．所有樣品，洗滌液和各種廢棄物都應(yīng)按傳染物處理。9．本試劑不同批號組分不得混用。10.如與英文說明書有異，以英文說明書為準。保存條件及有效期1．試劑盒保存：；2-8℃。2．有效期：6個月羊CD8分子ELISA試劑盒應(yīng)用雙抗體夾心法測定標本中羊CD8分子（CD8）水平。用純化的羊CD8分子（CD8）抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入CD8分子（CD8），再與HRP標記的CD8分子（CD8）抗體結(jié)合，形成抗體-抗原-酶標抗體復(fù)合物，經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍色，并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的CD8分子（CD8）呈正相關(guān)。用酶標儀在450nm波長下測定吸光度（OD值），通過標準曲線計算樣品中羊CD8分子（CD8）濃度。[詳細]
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2018-11-16 10:02
產(chǎn)品樣冊
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- 人黑色素ELISA Kit說明書
- 人黑色素ELISA Kit說明書[詳細]
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2014-08-21 00:00
安裝說明
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