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Millipore/CYT420 | <i>ChemiKine?</i> Pigment Epithelium Derived Factor, Sandwich ELISA/CYT420/1 kit
  • Millipore/CYT420 | <i>ChemiKine?</i> Pigment Epithelium Derived Factor, Sandwich ELISA/CYT420/1 kit

Millipore/CYT420 | ChemiKine? Pigment Epithelium Derived Factor, Sandwich ELISA/CYT420/1 kit

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貨號(hào): CYT420
品牌: Millipore
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    • Description
      CatalogueNumberCYT420
      BrandFamilyChemicon®
      TradeName
      • ChemiKine
      • Chemicon
      DescriptionChemiKine?PigmentEpitheliumDerivedFactor,SandwichELISA
      OverviewIntroduction

      PigmentEpithelium-DerivedFactor(PEDF)wasfirstidentifiedasafactorsecretedbyprimaryretinalpigmentepithelial(RPE)cellsthatpromotesthesurvivalofneuronsinvitroandinvivo.MolecularcharacterizationofPEDFshowsittobea50kDamemberoftheserpinfamilyofproteaseinhibitors1.AsecondmodeofactionofPEDFwasidentifiedwiththedemonstrationthatPEDFinducesapoptosisofmicrovascularendothelialcells,apropertythatmaintainstheavascularnatureoftheretina2.Asanumberofophthalmologicdisordersarecharacterizedbydysregulationoftheretinalinnervationandmicrovasculature,PEDFservesasanimportantregulatorofhumaneyedisease3.Inaddition,PEDFappearstohavemultifunctionalantitumoractivityinaneuroblastomamodel,asSchwanncell-derivedPEDFinducesadifferentiated,lessmalignantphenotypeinneuroblastomacells,promotesfurthergrowthandsurvivalofSchwanncells,andinhibitsangiogenesis4.

      ChemiKine?PigmentEpithelium-DerivedFactor(PEDF)kitisasandwichstripwell-typeenzymeimmunoassay(EIA),whichmeasuresPEDF.ThekitwillmeasurePEDFofvitreousfluidofhumanorigin.

      TestPrinciple

      WiththeChemiKine?PEDFassaysystem,amousemonoclonalantibodygeneratedagainsthumanPEDFiscoatedontoamicroplateandisusedtocapturePEDFfromasample.Asecond,biotinylatedPEDF-specificmousemonoclonalantibodydetectsthecapturedPEDF.Peroxidase-labeledstreptavidindetectsthebiotinylatedmousemonoclonalantibody.AfteradditionofthesubstrateandstopsolutiontheamountofPEDFisdetermined.ThestandardcurvedemonstratesadirectrelationshipbetweenOpticalDensity(OD)andPEDFconcentration:i.e.,thehighertheODthehigherthePEDFconcentrationinthesample.
      AlternateNames
      • PEDF
      MaterialsRequiredbutNotDelivered1.Multi-channelorrepeatingPipettes

      2.37oCincubator

      3.Platesealers

      4.Pipettors&tipscapableofaccuratelymeasuring10-1000mL

      5.GraduatedSEROlogicalpipettes

      6.96-wellmicroplatereaderwith450nmfilter

      7.Graphpaperformanualplottingofdata

      8.Polypropylenetesttubesforstandardandsampledilutions

      9.Mechanicalvortex

      10.One1or2litercontainer

      11.Urea
      ProductInformation
      Components
      • PEDFELISAPlate:(PartNo.61597)One96-WellImmunoplatePre-coatedwithMouseanti-HumanPEDFMonoclonalAntibody,sealedinafoilpouch.
      • WashBufferConcentrate:(PartNo.60245)One100mL(10X)bottleofConcentrate.
      • AssayDiluent:(PartNo.60240)One60mLbottle(ReadytoUse).
      • PEDFStandard(Human):(PartNo.61600)Onevialcontaining250ngrecombinantfulllengthhumanPEDF(Lyophilized).
      • BiotinylatedMouseanti-HumanPEDFMonoclonalAntibody:(PartNo.61598)One50μLvial.
      • Streptavidin,HRPconjugated:(PartNo.61599)One50μLvial.
      • TMB/ESolution:(PartNo.60096)One10mLbottleofaReadytoUsesolutionof3,3",5,5"-tetramethylbenzidineinaproprietarybufferwithenhancer.
      • StopSolution(PartNo.60193):One12mLbottleofanHClsolution(ReadytoUse).
      DetectionmethodColorimetric
      StorageandShippingInformation
      StorageConditionsMaintaintheunopenedkitat2-8°Cuntilexpirationdateindicatedonthelabel.Afteropeningthekitmaintaineachcomponentat2-8°Cuntilexpirationdateindicatedonthelabel.Afterreconstitution,maintainthePEDFStandardat-20oCforupto30days.Thestandardmaybefrozenandthawedthreetimeswithoutlossofimmunoreactivity.Donotstorereconstitutedstandardat4oC,aslossofimmunoreactivitywillresult.

      Precautions

      ·WashBufferandAssayDiluentcontainthimerosal.Thimerosalishighlytoxicbyinhalation,contactwithskinorifswallowed.ThimerosalisapossIBLemutagenandshouldbehandledaccordingly.

      ·Theinstructionsprovidedhavebeendesignedtooptimizethekit"sperformance.Deviationfromtheinstructionsmayresultinsuboptimalperformanceofthekitandthefailuretoproduceaccuratedata.
      Applications
      ApplicationThisChemiKinePigmentEpitheliumDerivedFactor,SandwichELISAisusedtomeasure&quantifyPEDFlevelsinNeuroscienceresearch.
      KeyApplications
      • ELISA
      ApplicationNotesChemiKine?PEDFkitisdesignedtomeasuretheamountofPEDFincellculturesupernatants,tissuehomogenatesandBIOLOGicalfluid(vitreous,serum,plasma,andserum-free)samplesofhumanorigin.Thereareenoughreagentsincludedinthiskitforone96-wellimmunoassayplate.Runningduplicatewellsforsamplesandstandardsisrecommended.

      ForResearchUseOnly.Notforuseindiagnosticprocedures.
      BiologicalInformation
      HostMouse
      SpeciesReactivity
      • Human
      AntibodyTypeMonoclonalAntibody
      AnalytesAvailable
      • PEDF
      EntrezGeneNumber
      GeneSymbol
      • SERPINF1
      • Serpin-F1
      • PEDF
      • PIG35
      • EPC-1
      UniProtNumber
      UniProtSummaryFUNCTION:SwissProt:P36955#Neurotrophicprotein;inducesextensiveneuronaldifferentiationinretinoblastomacells.Potentinhibitorofangiogenesis.AsitdoesnotundergotheS(stressed)toR(relaxed)conformationaltransitioncharacteristicofactiveserpins,itexhibitsnoserineproteaseinhibitoryactivity.
      SIZE:418aminoacids;46342Da
      SUBCELLULARLOCATION:Secreted.Melanosome.Note=EnrichedinstageImelanosomes.
      TISSUESPECIFICITY:Retinalpigmentepithelialcellsandbloodplasma.
      DEVELOPMENTALSTAGE:Expressedinquiescentcells.
      DOMAIN:SwissProt:P36955TheN-terminal(AA44-121)exhibitsneuriteoutgrowth-inducingactivity.TheC-terminalexposedloop(AA382-418)isessentialforserpinactivity.
      PTM:TheN-terminusisblocked.
      SIMILARITY:Belongstotheserpinfamily.
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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