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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1.单克隆抗体的优点:
(1)杂交瘤可以在体外“永久”地存活并传代,只要不发生细胞株的基因突变,就可以不断的生产高特异性、高均一性的抗体.
(2)可以用相对不纯的抗原,获得大量高度特异的、均一的抗体.
(3)由于可能得到“无限量”的均一性抗体,所以适用于以标记抗体为特点的免疫学分析方法,如IRMA和ELISA等.
(4)由于单克隆抗体的高特异性和单一生物学功能,可用于体内的放射免疫显像和免疫导向治疗.
2.单克隆抗体的局限性:
(1)单克隆抗体固定的亲和性和局限的生物活性限制了它的应用范围.由于单克隆抗体不能进行沉淀和凝集反应,所以很多检测方法不能用单克隆抗体完成.
(2)单克隆抗体的反应强度不如多克隆抗体.
(3)制备技术复杂,而且费时费工,所以单克隆抗体的价格也较高.
抗原有两个基本特性,即抗原性和免疫原性。有抗原性的物质不一定有免疫原性,所以由此引出半抗原和完全全抗原,半抗原必须经过经过一定的改造(偶联蛋白载体BSA,OVA或者HSA等大分子物质)方能成为完全。一般而言完全抗原分子量越大(大于10KDa),结构越复杂引起免疫反应的能力也就越强。
抗体就是能与特异性抗原结合的免疫球蛋白,抗体一般分为多克隆抗体和单克隆抗体,多克隆抗体能与抗原的多个表位结合。本篇主要讲述兔来源的多克隆抗体的生产步骤
多抗一般制备流程:完全抗原的准备→兔子的免疫→ 效价检测和终放→抗体亲和纯化→抗体的浓缩和保存。
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.
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。
其次,察看次目的蛋白的存在形式,有没有多聚体形式及变构形式;
最后,查看多家抗体公司的DATA,看看别人的WB做出来的条带的位置。
根据你说的,特异识别多个组织中的同样大小的条带,我觉得很可能就是你的目的蛋白。
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