Clone | Polyclonal |
ProductType | PrimaryAntibodies |
Units | 1ml |
Host | Rabbit |
Speciesreactivity | Human |
Application | immunoelectrophoresisPrecipitationassays |
BackgroundThedefinedantibodyreactivityisrestrictedtoFactorXIIIandFXIIIainplasmaandtoplateletFXIII.Astestedatthelevelofsensitivityofimmunoprecipitationtechniquesasingleprecipitinlineisobtainedinbidimensionalelectrophoresis,immunoelectrophoresisanddoublerADIalimmunodiffusion(Ouchterlony)whichshowsareactionoffullidentitywithAsubunitsofFXIII.NoreactionisobtainedwithFXIII-depletedplasmaandwithBsubunits.Inprecipitatingtechniquesaselectroimmunodiffusion,immunoelectrophoresisandsingleanddoubleradialimmunodiffusion(Mancini,Ouchterlony).ToprepareanadsorbentforimmunoaffinitypurificationofFXIII.Ifusedinmoresensitivetestproceduresorascatchingordetectionantibodyinsolidphaseimmunoassaysspecificitycontrolsshouldalwaysbeinclude.IncreaseinplasmaFXIIIhasbeenobservedintypeIVhyperlipoproteinaemiaandincombinedhyperlipodaemia.CongenitalandacquiredFXIIIdeficiencieshavealsobeenreported.Insomecasesofmyeloma,inhibitionofFXIIIactivitybytheparaproteinsmayoccur.
SourcePlasmaFXIIIisanactivetransglutaminaseuntilitisactivated(FXIIIa).ItconsistsoftwoAsubunitsjoinedasadimer,connectedtotwoBsubunits(A2B2).EachAsubunithasamolecularweightof75,000andeachBsubunitof80,000,makinganaggregatemolecularweightofthewholemoleculeof320,000.Normalplasmacontainsabout20mg/ml.PlateletFXIIIconsistsofonlyAsubunitssynthesizedinthemegakaryocytes.Itaccountsforabout50%ofthetotalbloodFXIIIactivitywithalinearrelationshipbetweenFXIIIlevelandplateletcounts.RAHu/FXIII-AreactswithbothplasmaandplateletFXIII.AntiserumraisedagainsttheBsubunit(RAHu/FXIII-S)reactsexclusivelywithnativeandactivatedFXIIIainplasma.FXIIIaresultsfromactivationofFXIIIbythrombinafterthereleaseoffibrinopeptidesAfromfibrinogen.HighlypurifiedFXIII-Aisusedfortheimmunization.Freund’scompleteadjuvantisusedinthefirststepoftheimmunizationprocedure.
ProductDelipidated,heatinactivated,lyophilized,stablewholeserum.Nopreservativeadded.TotalproteinandIgGconcentrationsintheantiserumarecomparabletothoseofpoolednormalrabbitserum.Noforeignproteinsadded.Reconstitutethelyophilizedantiserumbyadding1mlsteriledistilledwater.
SpecificityPrecipitatingpolyclonalRabbitantiSerumtoHumancoagulationfactorXIII-A.
SpeciesReactivity:TheantiSerumdoesnotcross-reactwithanyotherHumanplasmaproteinsastestedingel-diffusiontechniques.Inter-speciescross-reactivityisanormalfeatureofantibodiestoplasmaproteins,sincehomologousproteinsofdifferentspeciesfrequentlyshareantigenicdeterminants.Cross-reactivityofthisantiSerumhasnotbeentestedindetail.
ApplicationsPrecipitationassays.Inimmunoelectrophoresisinagarose-platesuse2μlhumanplasmaorequivalentagainst120μlantiserum.Indoubleradialimmunodiffusionusearosettearrangementwith10μlantiserumin3mmdiametercenterwelland2μlplasmasamples(neatandseriallydiluted)in2mmdiameterperipheralwells.Inelectroimmunodiffusiontheamountofantiserumrequiredintheagarosegelisusuallybetween1and2%dependingonthetestarrangement.
StorageThelyophilizedantiserumisshippedatambienttemperatureandmaybestoredat+4°C;prolongedstorageatorbelow-20°C.Dilutionsmaybepreparedbyaddingphosphatebufferedsaline(PBS,pH7.2).Repeatedthawingandfreezingshouldbeavoided.Ifaslightprecipitationoccursuponstorage,thisshouldberemovedbycentrifugation.Itwillnotaffecttheperformanceoftheantiserum.Dilutedantiserumshouldbestoredat+4°C,notrefrozen,andpreferablyusedthesameday.
CautionThisproductisintendedFORRESEARCHUSEONLY,andFORTESTSINVITRO,notforuseindiagnosticortherapeuticproceduresinvolvinghumansoranimals.Itmaycontainhazardousingredients. PleaserefertotheSafetyDataSheets(SDS)foradditionalinformationandproperhandlingprocedures.Disposeproductremaindersaccordingtolocalregulations.Thisdatasheetisasaccurateasreasonablyachievable,butExalphaBIOLOGicalsacceptsnoliABIlityforanyinaccuraciesoromissionsinthisinformation.
ProteinReference(s)
DatabaseName:UniProt
Accessionnumber:P00488
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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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