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Millipore/ECM335 | Human sICAM-1 ELISA/ECM335/1 kit
  • Millipore/ECM335 | Human sICAM-1 ELISA/ECM335/1 kit

Millipore/ECM335 | Human sICAM-1 ELISA/ECM335/1 kit

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貨號: ECM335
品牌: Millipore
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    • Description
      CatalogueNumberECM335
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionHumansICAM-1ELISA
      OverviewIntercellularAdhesionMolecule-1(ICAM-1)isamemberoftheimmunoglobulinsupergenefamily[Rothleinetal.,1988]andfunctionsasaligandforintegrinalphaLbeta2orLymphocyteFunction-AssociatedAntigen-1(LFA-1),amemberoftheleukocyteintegrinfamily[Marin&Springer1987]whichmediateslymphocyteadhesion.

      ICAM-1isasingle-chainglycoproteinwithapolypeptidecoreof55kDaexpressedonnon-hematopoieticcellsofmanylineagesincludingvascularendothelialcells,thymicepithelialcells,otherepithelialcellsandfibroblasts,andonhematopoieticcellssuchastissuemacrophages,mitogen-stimulatedT-lymphoblasts,germinalcenterB-cellsandthedendriticcellsfromtonsils,lymphnodesandPeyer"spatches.ICAM-1isinducIBLeonfibroblastsandendothelialcellsbytheinflammatorymediatorsIL-1,TNFandIFN-gamma.Itspresencecorrelateswithinfiltrationoflymphocytesintoinflammatorylesions[Dustinetal.,1989;Proberetal.,1986;Rothleinetal.,1986].ICAM-1seemstobetheinitialMarkerofinflammatoryreactionsandisexpressedpriortoandtoagreaterextentthanHLA-DR.

      TheroleofICAM-1asadiseasemarkerhasbeendemonstratedforanumberofdifferentpathologies,includingallergicrhinitis,allergiccontactdermatitis[Vejlsgaardetal.1989],gastrointestinalandbladdercancer,lymphoproliferativedisorders[Laletal.,1992;Rothleinetal.,1991],melanoma[Altomoneetal.,1992;Beckeretal.,1992;harningetal.,1991],HIV-1infection[Mostetal.,1993],malaria,tissueororgantransplantrejection[Adamsetal.,1989;Adamsetal.,1993],diabetesmellitus[Lampeteretal.,1992],glomerulonephritis,asthma[Wegneretal.,1990],rheumatoidarthritis[Popocniketal.,1990]andpsoriasis[Lisbyetal.,1989].

      Apanelof50serafromhealthyblooddonors(maleandfemale)wastestedforsICAM-1.ThedetectedsICAMlevelsrangedbetween129.9and297.4ng/mlwithameanof230.3ng/mlandastandarddeviationof47.4ng/ml.NormalsICAM-1levelsmayvarydependingontheserumcollectiveusedrangingupto400ng/mlorgreater.Dependinguponthestudypopulationrangesfrom219ng/mlto1050ng/mlhavebeenreportedintheliterature.

      AnalyticalSensitivityandDetectionLimits:

      Sensitivity:3.3ng/mL

      RangeofDetection:6.25-100ng/mL

      Intra-AssayVariation:4.1%

      Inter-AssayVariation:7.7%

      Recovery:98.6%average

      AssayTime:75minutes

      Crossreactivity:nointerferencefromsolubleTNF-R(60or80kDa),IL-8/NAP-1,TNF-alpha,TNF-beta,IFN-gamma,IFN-alpha2C,IFN-w,IL-6,IL-2R,E-selectin-1orL-selectin-1.

      TheICAM-1ELISAisanenzyme-linkedimmunosorbentassayforquantitativedetectionofsolubleIntercellularAdhesionMolecule-1incellculturesupernatants,serum,plasma,orotherBIOLOGicalfluids.TheICAM-1ELISAisforresearchuseonly.Notforuseindiagnosticprocedures.

      TestPrinciple:

      Ananti-ICAM-1monoclonalantibodyisadsorbedontomicrowells.ThepairofmonoclonalantibodiesuseinthisELISAassaydetectthesoluableformofICAM-1presentinserum,plasmaorotherbiologicalfluids.

      SolubleICAM-1presentinasampleorstandardthenbindstoantibodiesadsorbedtothemicrowells.Asecond,HRP-conjugatedmonoclonalanti-ICAM-1antibodyisaddedandbindstoICAM-1capturedbythefirstantibody.

      Unboundenzyme-conjugatedanti-ICAM-1isremovedwithawashstepandHRPsubstratesolutionisaddedtothewells.

      Anamountofcoloredproductisformed,proportionaltotheamountofsolubleICAM-1presentinthesample.Thereactionisterminatedbyadditionofacidandabsorbanceismeasuredat450nm.AstandardcurveispreparedfromfiveICAM-1standarddilutionsandtheICAM-1sampleconcentrationisdetermined.

      Application:

      TheICAM-1ELISAisanenzyme-linkedimmunosorbentassayforquantitativedetectionofsolubleIntercellularAdhesionMolecule-1incellculturesupernatants,serum,plasma,orotherbiologicalfluids.TheICAM-1ELISAisforresearchuseonly.Notforuseindiagnosticprocedures.
      AlternateNames
      • CD54
      MaterialsRequiredbutNotDelivered-5mLand10mLgraduatedPipettes

      -5μLto1000μLadjustablesinglechannelmicropipetteswith

      disposabletips

      -50μLto300μLadjustablemultichannelmicropipettewithdisposabletips

      -Multichannelmicropipettereservoir

      -Beakers,flasks,cylindersnecessaryforpreparationofreagents

      -Devicefordeliveryofwashsolution(multichannelwashbottleor

      automaticwashsystem)

      -MicrowellstripreadercapableofreADIngat450nm(620nmas

      optionalreferencewavelength)

      -Glass-distilledordeionizedwater

      -Statisticalcalculatorwithprogramtoperformlinearregression

      analysis.
      ProductInformation
      Components
      • ·1aluminumpouchwithMicrowellplatecoatedwithMonoclonalAntibody(murine)tohumanICAM-1
      • ·1vial(0.1mL)HRP-ConjugatedAnti-ICAM-1Monoclonal(murine)Antibody
      • ·2vials(0.5mLeach)100ng/mLsolubleICAM-1Standard
      • 1viallyophilizedcontrol(high).
      • 1viallyophilizedcontrol(low).
      • ·1bottle(50mL)WashBufferConcentrate20X(PBSwith1%Tween20)
      • ·1vial(5mL)AssayBufferConcentrate20X(PBSwith2%Tween20and10%BSA)
      • ·1bottle(12mL)SampleDiluent(bufferedproteinmatrix)
      • ·1vial(15mL)SubstrateSolution
      • ·1vial(15mL)StopSolution(1MPhosphoricAcid)
      • ·1MicrowellStripHolder
      • ·2AdhesivePlateCovers
      DetectionmethodChromogenic
      StorageandShippingInformation
      StorageConditionsStorekitreagents,exceptthecontrol,between2°and8°C.Storethelyophilizedcontrolat-20°C.Immediatelyafteruseremainingreagentsshouldbereturnedtocoldstorage(2°to8°C)withtheexceptionofreconstitutedcontrol(-20°C).Expirationdateofthekitisstatedonoutsidelabel.

      Precautions:

      ·Reagentsareintendedforresearchuseonlyandarenotforuseindiagnosticortherapeuticprocedures.

      ·Donotmixorsubstitutereagentswiththosefromotherlotsorothersources.

      ·Donotusekitreagentsbeyondexpirationdateonlabel.

      ·Donotexposekitreagentstostronglightduringstorageor

      incubation.

      ·Rubberordisposablelatexglovesshouldbewornwhilehandlingkitreagentsorspecimens.

      ·Somereagentscontainthimerosalaspreservative,whichishighlytoxicbyinhalation,ingestion,orcontactwithskin.Thimerosalisapossiblemutagenandshouldbehandledaccordingly.

      ·Avoidcontactofsubstratesolutionswithoxidizingagentsandmetal.

      ·Inordertoavoidmicrobialcontaminationorcross-contaminationofreagentsorspecimenswhichmayinvalidatethetestusedisposablepipettetipsand/orpipettes.

      ·Useclean,dedicatedreagenttraysfordispensingtheconjugateandsubstratereagents.

      ·Exposuretoacidswillinactivatetheconjugate.

      ·Glass-distilledwaterordeionizedwatermustbeusedforreagent

      preparation.

      ·Substratesolutionsmustbeatroomtemperaturepriortouse.

      ·Sinceexactconditionsmayvaryfromassaytoassay,astandard

      curvemustbeestablishedforeveryrun.

      ·Bacterialorfungalcontaminationofeitherscreensamplesorreagentsorcross-contaminationbetweenreagentsmaycauseerroneousresults.

      ·Disposablepipettetips,flasksorglasswarearepreferred.Reusableglasswaremustbewashedandthoroughlyrinsedofalldetergentsbeforeuse.

      ·Improperorinsufficientwashingatanystageoftheprocedurewillresultineitherfalsepositiveorfalsenegativeresults.CompletelyemptywellsbeforedispensingfreshWashBuffer,fillwithWashBufferasindicatedforeachwashcycleanddonotallowwellstosituncoveredordryforextendedperiods.

      ·Theuseofradioimmunotherapyhassignificantlyincreasedthenumberofpatientswithhumananti-mouseIgGantibody(HAMA).HAMAmayinterferewithassaysutilizingmurinemonoclonalantibodiesleadingtobothfalsepositiveandfalsenegativeresults.HAMAinterferencecanbereducedbyaddingmurineimmunoglobulins(serum,asciticfluid,ormonoclonalantibodiesofirrelevantspecificity)totheSampleDiluent.
      Applications
      ApplicationThisHumansICAM-1ELISAisusedtomeasure&quantifysICAM-1levelsinInflammation&Immunology&Neuroscienceresearch.
      KeyApplications
      • ELISA
      BiologicalInformation
      SpeciesReactivity
      • Human
      AnalytesAvailable
      • sICAM-1
      EntrezGeneNumber
      EntrezGeneSummaryICAM1(CD54)istypicallyexpressedonendothelialcellsandcellsoftheimmunesystem.ICAM1bindstointegrinsoftypeCD11a/CD18,orCD11b/CD18.ICAM1isalsoexploitedbyRhinovirusasareceptor.
      GeneSymbol
      • ICAM1
      • P3.58
      • BB2
      • CD54
      • ICAM-1
      UniProtNumber
      UniProtSummaryFUNCTION:SwissProt:P05362#ICAMproteinsareligandsfortheleukocyteadhesionproteinLFA-1(integrinalpha-L/beta-2).Incaseofrhinovirusinfectionactsasacellularreceptorforthevirus.Alsoactsasreceptorforsomecoxsackieviruses.
      SIZE:532aminoacids;57825Da
      SUBUNIT:Interactswithhumanherpesvirus8MIR2protein(Probable).
      SUBCELLULARLOCATION:Membrane;Single-passtypeImembraneprotein.
      PTM:Monoubiquitinated,whichispromotedbyMARCH9andleadstoendocytosis.
      SIMILARITY:SwissProt:P05362##Belongstotheimmunoglobulinsuperfamily.ICAMfamily.&Contains5Ig-likeC2-type(immunoglobulin-like)domains.
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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