BackgroundTheCD3epitopeisexpressedontheepsilonchainoftheCD3/Tcellantigenreceptor(TcR)complex.CD3ispresenton65-85%ofthymocytesandhasamitogeniceffectonperipheralbloodTcells.CD3indentifieshumanTcellsexpressingthe22-28,000M.W.surfaceantigen.CD4identifieshumanhelper/inducerTcellsexpressingthe60,000M.W.surfaceantigen(HLAclassIIreactive).CD4ispresentinlowdensityonmonocytes.IdentificationofCD8onhumancytotoxic/suppressorTcellsexpressingthe32and43,000M.W.surfaceantigens.

Synonyms:CD3/4/8

Source

Immunogen:CD4=DerivedfromthehybridizationofmouseNS-1myelomacellswithspleencellsfromBALB/cmiceimmunizedwithhumanperherialbloodTlymphocytes.CD3=DerivedfromthehybridizationofmouseNS-1myelomacelsswithspleencellsfromBALB/cmiceimmunizedwithhumanthymocytes.CD8=DerivedfromthehybridizationofmouseNS-1myelomacellswithspleencellsfromBALB/cmiceimmunizedwithhumanperherialbloodTlymphocytes.

Product

ProductForm:RPE-Cy-5

Formulation:Providedassterilefilteredsolutioninphosphatebufferedsalinewith0.08%sodiumazideand0.2%carrierprotein

PurificationMethod:AffinityChromatography

Concentration:Titeredforflowcytometry

ApplicationsPBMC:Add15µlof3-ColorTMantibodyreagent/10^6PBMCin100µlPBS.Mixgentlyandincubatefor15minutesat2°to8°C.WashtwicewithPBS.WashtwicewithPBSorfixwith0.5%v/vofparaformaldehydeinPBSandanalyze.WHOLEBLOOD:Add15µlofExalphaBIOLOGical"s3-ColorTMantibodyreagentMAB/100µlofwholeblood.Mixgentlyandincubatefor15minutesatroomtemperature20°C.Lysethewholeblood.WashtwicewithPBSandanalyzeorfixwith0.5%v/vofparaformaldehydeinPBSandanalyze.Seeinstrumentmanufacturer’sinstructionsforLysedWholeBloodandImmunofluorescenceanalysiswithaflowcytometerormicroscope.

FunctionalAnalysis:FlowCytometryStaining

StorageProductshouldbestoredat4-8ºC.DONOTFREEZE

ProductStABIlity:Reagentsarestablefortheperiodshownontheviallabelwhenstoredproperly

ShippingConditions:RoomTemperature

CautionThisproductisintendedFORRESEARCHUSEONLY,andFORTESTSINVITRO,notforuseindiagnosticortherapeuticproceduresinvolvinghumansoranimals.Itmaycontainhazardousingredients. PleaserefertotheSafetyDataSheets(SDS)foradditionalinformationandproperhandlingprocedures.Disposeproductremaindersaccordingtolocalregulations.Thisdatasheetisasaccurateasreasonablyachievable,butNordic-Mubio​acceptsnoliabilityforanyinaccuraciesoromissionsinthisinformation.

References1.OrfaoA,GonzálezdeBuitragoJMLacitometríadeflujoenellaboratorioclínico.SociedadespañoladebioquímicaclínicaypatologíaMolecular1995.2.Stetler-StevensonM.Flowcytometryanalysisoflymphomasandlymphoproliferativedisorders.SeminHematol2001Apr;38(2):111-23.3.MenéndezP,etal.Comparisonbetweenalyse-and-then-washmethodandalyse-non-washtechniquefortheenumerationofCD34+hematopoieticProgenitorcells.Cytometry(Comm.Clin.Cytometry)34:264-271(1998)4.GratamaJW,MenéndezP,KraanJ,OrfaoA.LossofCD34+hematopoieticprogenitorcellsduetowashingcanbereducedbytheuseoffixative-freeerytrocytelysingreagents.JImmunol.Methods239:13-23(2000)5.ProtectionofLaboratoryWorkersfromoccupationallyacquiredinfections.Secondedition;approvedguideline(2001).VillanovaPA:NationalCommitteeforClinicalLaboratoryStandards;DocumentM29-A2.6.Proceduresforthecollectionofdiagnosticbloodspecimensbyvenipuncture-approvedstandard;Fifthedition(2003).WaynePA:NationalCommitteeforClinicalLaboratoryStandards;DocumentH3-A5.7.Clinicalapplicationsofflowcytometry:Qualityassuranceandimmunophenotypingoflymphocytes;approvedguideline(1998).WaynePA:NationalCommitteeforClinicalLaboratoryStandards;DocumentH42-A.8.LimaMetal.TCRαβ+/CD4+LargeGranularLymphocytosis.AnewClonalT-CellLymphoprolipherativeDisorder.AmericanJournalofPathology,163(2):763-771(2003)9.GorczyzaW.etal.AnapproachtodiagnosisofT-celllymphoprolipherativedisordersbyflowcytometry.Cytometry(Clinicalcytometry)50:177-190(2002)10.BraylanRC,OrfaoA,BorowitzMJ,DavisBH.Optimalnumberofreagentsrequiredtoevaluatehematolymphoidneoplasias:resultsofaninternationalconsensusmeeting.Cytometry46:23-7(2001)11.JenningsCD,FoonKA.Recentadvancesinflowcytometry:applicationtothediagnosisofhematologicmalignancy.Blood90(8):2863-2892(1997)12.Reichertetal.LymphocytesubsetreferencerangesinadultCaucasians.ClinImmunolImmunopathol60:190-208(1991)13.PrinceHKetal.InfluenceofracialbackgroundonthedistributionofT-cellsubsetsandLeu-11positivelymphocytesinhealthyblooddonors.DiagnImmunol.3:33-39(1985)14.KotyloPKetal.Referencerangesforlymphocytesubsetsinpediatricpatients.AmJClinPathol100:111-5(1993)Cy-5PortionsofthisproductismanufacturedunderlicensefromCarnegieMellonUniversity,U.S.PatentNumber5,268,486.

ProteinReference(s)

DatabaseName:UniProt

Accessionnumber:P07766,P01730,P01732

SpeciesAccession:Human

SafetyDatasheet(s)forthisproduct:NM_SodiumAzide/wp-content/uploads/SDS/AntibodySDSwithSodiumAzideNoridic-MUbio.pdf