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

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貨號: ECM335

品牌: Millipore

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Description
CatalogueNumber	ECM335
BrandFamily	Chemicon®
TradeName	Chemicon
Description	HumansICAM-1ELISA
Overview	IntercellularAdhesionMolecule-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
Detectionmethod	Chromogenic

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

Detectionmethod

Chromogenic

StorageandShippingInformation
StorageConditions	Storekitreagents,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.

StorageandShippingInformation

StorageConditions

Storekitreagents,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
Application	ThisHumansICAM-1ELISAisusedtomeasure&quantifysICAM-1levelsinInflammation&Immunology&Neuroscienceresearch.
KeyApplications	ELISA

BiologicalInformation
SpeciesReactivity	Human
AnalytesAvailable	sICAM-1
EntrezGeneNumber	NM_000201.1
EntrezGeneSummary	ICAM1(CD54)istypicallyexpressedonendothelialcellsandcellsoftheimmunesystem.ICAM1bindstointegrinsoftypeCD11a/CD18,orCD11b/CD18.ICAM1isalsoexploitedbyRhinovirusasareceptor.
GeneSymbol	ICAM1 P3.58 BB2 CD54 ICAM-1
UniProtNumber	P05362
UniProtSummary	FUNCTION: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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Millipore/ECM335 | Human sICAM-1 ELISA/ECM335/1 kit

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