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Millipore/ES022-100ML | TMB/E Ultra Sensitive, Blue, Horseradish Peroxidase Substrate (soluble)/ES022-100ML/100 mL
  • Millipore/ES022-100ML | TMB/E Ultra Sensitive, Blue, Horseradish Peroxidase Substrate (soluble)/ES022-100ML/100 mL

Millipore/ES022-100ML | TMB/E Ultra Sensitive, Blue, Horseradish Peroxidase Substrate (soluble)/ES022-100ML/100 mL

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货号: ES022-100ML
品牌: Millipore
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    • Description
      CatalogueNumberES022-100ML
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionTMB/EUltraSensitive,Blue,HorserADIshPeroxidaseSubstrate(soluble)
      OverviewTMB,(3,3",5,5"-tetramethybenzidine)hasbeenshowntobeasafe-sensitivesubstratefortheassayofHRP.Initially,inthepresenceofHRPandhydrogenperoxide,aone-electronoxidationproductisformed.Thiscompound,acationfreeradical,isblueincolorwithanadsorptionmaximumof653nm.FurtherreactionwithHRP/H2O2oracidificationoftheradicalwithacidyieldsthediimineterminaloxidationproductadsorbinglightat450nm.Theextinctioncoefficientoftheradical(E653nm=3.9x104mol-1cm-1)anddiimine(E450nm=5.9x104mol-1cm-1)providearemarkablysensitivesystemfortheassayofHRPandHRPlabeledprobes.ES022isasinglecomponentreagentstableatroomtemperatureandnotsensitivetonormallaboratorylight.ItisoptimizedwithrespecttoTMBandhydrogenperoxideconcentrationsandyieldsalinearresponsewiththeconcentrationsofHRPusuallyemployedinimmunologicalassays.
      ProductInformation
      PresentationREAGENTPROVIDED:

      LiquidsolutioncontainingTMB,2.08μMolL(-1)andHydrogenPeroxide,citricacidbufferatpH3.3.Alsocontainsnon-toxicproprietarystABIlizers.

      MATERIALSREQUIRED:ReagentsRequiredforStoppingtheReaction,ButNotProvided

      A.0.3MolLsulphuricacidforstoppingthereactionandpreservingbluechromogen.

      Note:Reagentgradewatermustcontainlessthan10(-7)Mol/LofironorcoppersaltsotherwiseunreactedTMBwillbeconvertednon-enzymaticallytothediimine.
      ColorCodeYellow
      StorageandShippingInformation
      StorageConditionsMaintainatroomtemperatureforuptosixmonths.Forlongtermstorage,uptotwelvemonths,maintainat2-8°C.Protectfromexposuretodirectsunlight.Discardifsolutionisblueorturbid.

      Warmtoroomtemperaturepriortouse.

      REAGENTPROVIDED:LiquidsolutioncontainingTMB,2.08mMolL(-1)andHydrogenPeroxide,citricacidbufferatpH3.3.Alsocontainsnon-toxicproprietarystabilizers.
      Applications
      Application100mlTMB/EUltraSensitive,Blue,HorseradishPeroxidaseSubstrate(soluble)forELISA&Immunohistochemistry.
      KeyApplications
      • ELISA
      • Immunohistochemistry
      ApplicationNotesMETHODSYNOPSIS:


      AftercompletionofanalytebindingtoasolidphaseandreactionwithaHRPlabeledprobe,ES022isadded.OxidationofTMBproducesabluereactionproductthatismeasuredat650nm.Thecolorformationasafunctionoftimecanberecordedorthereactionstoppedwithsodiumfluorideafterafixedinterval.Increasedsensitivitycanbeachievedbyconvertingtheblueradicaltothediiminebyadditionofacid.Theresultingyellowcolorismeasuredimmediatelyat450nm.


      SAMPLEPROCEDURE:


      1.Completeallrequiredincubationswithantibodies,probes,HRPlabeledreagents.


      2.Washplateatleast4timeswithPBSorTrisbufferedsalinecontaining0.1%Tween-20.


      3.Afterthefinalwash,shakeandblotallresidualbufferfromplatewells.


      4.Add0.1mLofES022(TMBESolution)toappropriatewellsandincubate5-30minutes.


      Note:ThereactiontimewilldependupontheactivityoftheHRPprobe.Ifcolordevelopstoobriskly,zeroorderkineticswillnotprevail.Dilutionoftheprobe,antibodyorHRPlabeledreagentmayberequired.


      5.Thereactioncanbemonitoredasafunctionoftimeforkineticassaysorstoppedwith0.1mLof0.3Mol/Lofsulphuricacidor0.1%sodiumfluorideandreadat450nm.


      6.Iftheproceduredemandsconversiontotheyellowdiimine,add0.1mLofeitheraciddescribedinthereagentsectionaboveandrecordtheabsorbancewithin5minutes.


      Variationsoftime,reagentvolumeandtemperaturerequirestandardizationbytheuser.
      BIOLOGicalInformation
      PhysicochemicalInformation
      QuantitationWavelength450nm
      Dimensions
      MaterialsInformation
      MaterialsInformation
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