| ProductType | Three-ColorReagents(FITC/RPE/Cy5RPE) |
| Units | 50Tests |
| Host | Mouse |
| Speciesreactivity | Human |
| Application | FlowCytometryImmunofluorescence |
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-Mubioacceptsnoliabilityforanyinaccuraciesoromissionsinthisinformation.
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
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制备抗体都需要进行纯化处理
又由于自然存在的抗原大都存在多个抗原表位,会刺激机体产生多种针对同一抗原的不同抗原表位相应的不同抗体.
清冷冻干燥后保存。
一般来说多克隆的阳性率高一些，但出现假阳性的比例也高一些。
抗体的特异性鉴定 抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高，它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线，计算各自的结合率，求出各自在 IC50时的浓度，并按下列公式计算交叉反应率。 如果所用抗原浓度IC50浓度为pg/管，而一些近似抗原物质的IC50浓度几乎是无穷大时，表示 这一抗血清与其他抗原物质的交叉反应率近似为 0，即该血清的特异性较好。
抗体的亲和力 是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小、抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键、疏水键、侧链相反电荷基因的库仑力、范德华力和空间斥力。亲和力常以亲和常数K表示，K的单位是L/mol，通常K的范围在 108 ～1010 /mol，也有多达 1014 /mol。抗体亲和力的测定对抗体的筛选，确定抗体的用途，验证抗体的均一性等均有重要意义。向左转|向右转
