Human HMW Adiponectin Quantikine ELISA Kit

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• Human HMW Adiponectin Quantikine ELISA Kit

  HumanHMWAdiponectinQuantikineELISAKit[详细]

  2018-10-01 10:01

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• Human uPAR Quantikine ELISA Kit

  HumanuPARQuantikineELISAKit[详细]

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• Human VEGF Immunoassay Quantikine ELISA

  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领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领.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领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.[详细]

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• Human interleukin-17(IL-17) ELISA kit

  Human interleukin-17(IL-17) ELISA kit[详细]

  2015-10-09 00:00

  实验操作

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• 人(Human)脂联素（Adiponectin）ELISA 检测试剂盒

  人(Human)脂联素（Adiponectin）ELISA 检测试剂盒[详细]

  2015-01-22 00:00

  选购指南

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• 人（Human）白介素1α（IL-1α）ELISA Kit使用说明书

  人原钙黏素1（PCDH1）ELISAKit人白介素2受体（IL-2R）ELISAKit人皮肤T细胞虏获趋化因子（CTACK/CCL27）ELISAKit人胸肾表达趋化因子（BRAK/CXCL14）ELISAKit人B-淋巴细胞趋化因子1（BLC-1/CXCL13）ELISAKit人结缔组织活化肽Ⅲ（CTAPⅢ）ELISAKit人巨噬细胞集落刺激因子受体（M-CSFR）ELISAKit人免疫球蛋白AFc段受体Ⅰ（FcαRⅠ/CD89）ELISAKit人免疫球蛋白EFc段受体Ⅱ（FcεRⅡ/CD23）ELISAKit人免疫球蛋白GFc段受体Ⅲ（FcγRⅢ/CD16）ELISAKit人免疫球蛋白GFc段受体Ⅱ（FcγRⅡ/CD32）ELISAKit人免疫球蛋白GFc段受体Ⅰ（FcγRⅠ/CD64）ELISAKitGFc段受体Ⅰ（FcγRⅠ/CD64）ELISAKit人免疫球蛋白GFc段受体Ⅲ（FcγRⅢ/CD16）ELISAKit人免疫球蛋白GFc段受体Ⅱ（FcγRⅡ/CD32）ELISAKit人免疫球蛋白GFc段受体Ⅰ（FcγRⅠ/CD64）ELISAKit人粒细胞趋化蛋白-2（GCP-2/CXCL6）ELISAKit人ω干扰素（IFN-ω）ELISAKit人糖基化依赖的细胞黏附分子（GlyCAM-1）ELISAKit人干扰素调节因子（IRF）ELISAKit人淋巴毒素β（LTB）ELISAKit人淋巴毒素α（LTA）ELISAKit人CC趋化因子受体1（CCR1）ELISAKit人CX3C趋化因子受体1（CX3CR1）ELISAKit人肺部活化调节趋化因子（PARC/CCL18）ELISAKit人黏膜地址素细胞黏附分子（MAdCAM-1）ELISAKit人β干扰素（IFN-β/IFNB）ELISAKit人可溶性CD38（sCD38）ELISAKit人可溶性CD21（CR2/sCD21）ELISAKit人可溶性瘦素受体（sLR）ELISAKit人Toll样受体9（TLR-9/CD289）ELISAkit人转化生长因子β2（TGFβ2）ELISAKit人单核细胞趋化蛋白4（MCP-4/CCL13）ELISAKit人白三烯D4（LTD4）ELISAKit人N钙黏蛋白/神经钙黏蛋白（N-Cad）ELISAKit人肝素结合性表皮生长因子（HB-EGF）ELISAKit人红细胞刺激因子（ESF）ELISAKit人肿瘤坏死因子相关激活诱导因子（TRANCE）ELISAKit人生长激素释放因子（GH-RF）ELISAKit人巨噬细胞趋化因子（MCF）ELISAKit人α/β干扰素受体（IFN-α/βR）ELISAKit人B细胞生长因子（BCGF）ELISAKit人B细胞分化因子（BCDF）ELISAKit人上皮细胞粘附分子（Ep-CAM/CD362）ELISAKit人可溶性粘附分子（Sam）ELISAKit人巨噬细胞替代激活相关化学因子1（AmAC-1）ELISAKit人可溶性血管内皮生长因子受体2（VEGFR-2/sFLK-1）ELISAKit人胸腺基质淋巴细胞生成素（TSLP）ELISAKit人穿孔素/成孔蛋白（PF/PFP）ELISAKit人多效生长因子（PTN）ELISAKit人可溶性CD28（sCD28）ELISAKit人淋巴细胞因子ELISAKit人胸腺活化调节趋化因子（TARC/CCL17）ELISAKit人神经细胞粘附分子配体1（NCAM-L1/CD171）ELISAKit人神经保护因子（CVNPF）ELISAKit人可溶性肿瘤坏死因子α受体（sTNFαR）ELISAKit人可溶性细胞因子受体（sCKR）ELISAKit人可溶性凋亡相关因子配体（sFASL）ELISAKit人细胞凋亡YZ因子（IAP）ELISAKit人集落刺激因子（CSF）ELISAKit人γ干扰素诱导单核细胞因子（MIGF/CXCL9）ELISAKit人干扰素诱导T细胞趋化因子（ITAC/CXCL11）ELISAKit人CD14分子（CDl4）ELISAKit人凋亡诱导因子（AIF）ELISAKit人白细胞共同抗原（LCA/CD45）ELISAKit人CD4分子（CD4）ELISAKit人P钙黏蛋白/胎盘钙黏蛋白（P-cad）ELISAKit人角化细胞生长因子（KGF）ELISAKit人血小板衍生生长因子BB（PDGF-BB）ELISAKit人CXC趋化因子配体16（CXCL16）ELISAKit人CXC趋化因子受体3（CXCR3）ELISAKit人γ干扰素诱导蛋白16/p16（IFI16/p16）ELISAKit人基质细胞衍生因子1a（SDF-1a/CXCL12）ELISAKit人淋巴细胞趋化因子（Lptn/LTN/XCL1）ELISAKit人α干扰素（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人巨噬细胞移动YZ因子（MIF）ELISAKit人组织因子途径YZ物（TFPI）ELISAKit人干扰素诱导蛋白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人表皮生长因子（EGF）ELISAKit人碱性成纤维细胞生长因子（bFGF）ELISAKit人巨噬细胞炎性蛋白5（MIP-5）ELISAKit人可溶性E选择素（sE-selectin）ELISAKit人可溶性细胞间粘附分子1（sICAM-1）ELISAKit人细胞间粘附分子2（ICAM-2/CD102）ELISAKit人细胞间粘附分子3（ICAM-3/CD50）ELISAKit人结缔组织生长因子（CTGF）ELISAKit人白介素18（IL-18）ELISAKit人粘膜相关上皮趋化因子（MEC/CCL28）ELISAKit人粘膜相关上皮趋化因子（MEC/CCL28）ELISAKit人B细胞活化因子受体（BAFF-R）ELISAKit人血管内皮细胞生长因子受体3（VEGFR-3/Flt-4）ELISAKit人血管内皮细胞生长因子受体1（VEGFR-1/Flt1）ELISAKit人血管内皮细胞生长因子D（VEGF-D）ELISAKit[详细]

  2024-09-30 09:18

  产品样册

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• Monkey ELISA Kit

  MonkeyELISAKit[详细]

  2024-09-28 07:04

  产品样册

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• 上海易利为您提供Human GST elisa kit 英文说明书

  本公司为ELISA试剂盒供应商，价格公正，售前，售中，售后全程为您服务。提供免费代测服务，一周内出结果，保证质量欢迎来电咨询!许经理021-6051540918217407991[详细]

  2018-10-01 10:00

  产品样册

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• 人凝血酶ELISA Kit

  人凝血酶ELISA Kit[详细]

  2014-08-21 00:00

  安装说明

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• Mouse CaSR ELISA kit

  Mouse CaSR ELISA kit[详细]

  2015-10-09 00:00

  操作手册

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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领specimensandreagents.Microbialcontaminationofreagentsorspecimensmaygivefalseresults.lHand领首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:[详细]

  2018-09-22 10:00

  产品样册

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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[详细]

  2018-10-31 10:00

  产品样册

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• Human 8-OHDG elisa试剂盒

  Human 8-OHDG elisa试剂盒[详细]

  2024-09-16 18:47

  期刊论文

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• 人ELISA试剂盒,TPA ELISA Kit

  96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原（TPA）ELISA试剂盒相关产品：96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原（TPA）ELISA试剂盒IL-10,小鼠白介素-10Elisa试剂盒磺胺二甲基恶唑,标准品JLJL200712人Elisa试剂盒,人ProteinCElisa试剂盒，HumanProteinCELISA试剂盒CAS号:108-69-0,3,5-二,98%人Elisa试剂盒,人CD30Elisa试剂盒，HumanClusterofdifferentiation30,CD30ELISA试剂盒CAS:893-36-7,盐酸-L-白氨酰-2-萘胺/L-亮氨酰-2-萘胺盐酸盐/L-白氨酰-β-萘胺盐酸盐/盐酸-L-亮氨酰-2-萘胺/L（+）-亮氨酰-2-萘基盐酸氨/L-Leucyl-2-naphthylamidehydrochloride,BR,98%,1克,避光,-20℃96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原（TPA）ELISA试剂盒Humanbacterialvaginosis,BVELISA试剂盒人（BV）kit说明书,细菌性阴道病Elisa试剂盒CXCR3ELISAKit,大鼠CXC趋化因子受体3Elisa检测试剂盒蒙花苷,标准品,含量测定,20mg,常温,避光PorcineapoproteinA1,apo-A1ELISA试剂盒猪（apo-A1）kit说明书,载脂蛋白A1Elisa试剂盒大鼠淋巴细胞因子ELISA试剂盒HumanhepatitisBvirusXinteractingprotein,HBXIPELISAKit人异常凝血酶原（APT）ELISA试剂盒HumanAbnormalprothrombin,APTELISA试剂盒草乌甲素,标准品,含量测定,50mg,常温,避光96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原（TPA）ELISA试剂盒GRP1957,营养肉汤,供一般细菌培养、转种和增菌用,250g小鼠生长激素释放多肽（GHRP）ELISA试剂盒HumanMotilin,MTLELISAKitCAS号:4767-3-7,2,2-双（羟甲基）丙酸,98%[详细]

  2018-10-23 10:31

  产品样册

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• [ELISA]CD8分子（CD8）ELISA Kit

  羊CD8分子（CD8）试剂盒使用说明书本试剂盒仅供研究使用。检测范围：96T7IU/ml-240IU/ml试剂盒组成130倍浓缩洗涤液20ml×1瓶7终止液6ml×1瓶2酶标试剂6ml×1瓶8标准品（400IU/ml）0.5ml×1瓶3酶标包被板12孔×8条9标准品稀释液1.5ml×1瓶4样品稀释液6ml×1瓶10说明书1份5显色剂A液6ml×1瓶11封板膜2张6显色剂B液6ml×1/瓶12密封袋1个标本要求1．标本采集后尽早进行提取，提取按相关文献进行，提取后应尽快进行实验。若不能马上进行试验，可将标本放于-20℃保存，但应避免反复冻融2．不能检测含NaN3的样品，因NaN3YZ辣根过氧化物酶的（HRP）活性。羊CD8分子ELISA试剂盒用于测定羊血清、血浆及相关液体样本中CD8分子（CD8）含量。操作步骤1.标准品的稀释：本试剂盒提供原倍标准品一支，用户可按照下列图表在小试管中进行稀释。200IU/ml5号标准品150μl的原倍标准品加入150μl标准品稀释液100IU/ml4号标准品150μl的5号标准品加入150μl标准品稀释液50IU/ml3号标准品150μl的4号标准品加入150μl标准品稀释液25IU/ml2号标准品150μl的3号标准品加入150μl标准品稀释液12.5IU/ml1号标准品150μl的2号标准品加入150μl标准品稀释液2.加样：分别设空白孔（空白对照孔不加样品及酶标试剂，其余各步操作相同）、标准孔、待测样品孔。在酶标包被板上标准品准确加样50μl，待测样品孔中先加样品稀释液40μl，然后再加待测样品10μl（样品Z终稀释度为5倍）。加样将样品加于酶标板孔底部，尽量不触及孔壁，轻轻晃动混匀。3.温育：用封板膜封板后置37℃温育30分钟。4.配液：将30倍浓缩洗涤液用蒸馏水30倍稀释后备用5.洗涤：小心揭掉封板膜，弃去液体，甩干，每孔加满洗涤液，静置30秒后弃去，如此重复5次，拍干。6.加酶：每孔加入酶标试剂50μl，空白孔除外。7.温育：操作同3。8.洗涤：操作同5。9.显色：每孔先加入显色剂A50μl，再加入显色剂B50μl，轻轻震荡混匀，37℃避光显色15分钟.10.终止：每孔加终止液50μl，终止反应（此时蓝色立转黄色）。11.测定：以空白空调零，450nm波长依序测量各孔的吸光度（OD值）。测定应在加终止液后15分钟以内进行。羊CD8分子ELISA试剂盒注意事项1．试剂盒从冷藏环境中取出应在室温平衡15-30分钟后方可使用，酶标包被板开封后如未用完，板条应装入密封袋中保存。2．浓洗涤液可能会有结晶析出，稀释时可在水浴中加温助溶，洗涤时不影响结果。3．各步加样均应使用加样器，并经常校对其准确性，以避免试验误差。一次加样时间**控制在5分钟内，如标本数量多，推荐使用排枪加样。4．请每次测定的同时做标准曲线，**做复孔。如标本中待测物质含量过高（样本OD值大于标准品孔**孔的OD值），请先用样品稀释液稀释一定倍数（n倍）后再测定，计算时请Z后乘以总稀释倍数（×n×5）。5．封板膜只限一次性使用，以避免交叉污染。6．底物请避光保存。7．严格按照说明书的操作进行，试验结果判定必须以酶标仪读数为准.8．所有样品，洗涤液和各种废弃物都应按传染物处理。9．本试剂不同批号组分不得混用。10.如与英文说明书有异，以英文说明书为准。保存条件及有效期1．试剂盒保存：；2-8℃。2．有效期：6个月羊CD8分子ELISA试剂盒应用双抗体夹心法测定标本中羊CD8分子（CD8）水平。用纯化的羊CD8分子（CD8）抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入CD8分子（CD8），再与HRP标记的CD8分子（CD8）抗体结合，形成抗体-抗原-酶标抗体复合物，经过彻底洗涤后加底物TMB显色。TMB在HRP酶的催化下转化成蓝色，并在酸的作用下转化成Z终的黄色。颜色的深浅和样品中的CD8分子（CD8）呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），通过标准曲线计算样品中羊CD8分子（CD8）浓度。[详细]

  2018-11-16 10:02

  产品样册

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• 人黑色素ELISA Kit说明书

  人黑色素ELISA Kit说明书[详细]

  2014-08-21 00:00

  安装说明

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