| 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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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。
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。
1取组织
（1） 灌注固定：暴露心脏,将注射针头插入左心室,剪开右心耳,快速灌注PBS,待右心耳处流出澄清液体,换4%多聚甲醛灌注到大鼠四肢强直为止,快速取脑和肾。
（2） 固定：将组织置于4%多聚甲醛中固定15小时
（3） 脱水：将组织放入30% 蔗糖溶液中直至下沉
（4） 切片：脑组织冰冻切片20微米，37度烘干2h以上；
（5）将切片放置－80度保存，随用随取。
2免疫单标记
（1）做抗原修复（0.4g柠檬酸、3g柠檬酸钠、1000ml水配制成溶液，放到水浴箱中90~94℃保存15min）,冷却后， 用1*PBS反复洗标本10min，吸去PBS，同时向标本加渗透液（配0.4%trition液，然后将BSA加入到配好的trition液中，配成1％浓度）100ul，常温下湿盒反应1h。
（2） 吸去渗透液，同时向标本加block液（5％封闭用正常山羊血清）100ul，常温下湿盒反应5h。
（3） 吸去block液，同时向标本加入1：50稀释的羊抗鼠一抗100ul，37度湿盒反应过夜。
（4）吸去一抗，并回收，用1*PBS洗液反复洗15min。
（5）快速从冰箱中取出驴抗羊的二抗，并立即用锡纸包裹，注入到一个新的以锡纸包裹的tube管中并用PBS以1：200稀释。
（6）进入暗室，吸去PBS，同时加入二抗100ul，37度湿盒反应2h,封片。
（7）放置到荧光显微镜下观察。
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.
