| Isotype | IgG |
| ProductType | BlockingPeptide |
| Units | 50µg |
| Host | Rabbit |
| Speciesreactivity | Human |
| Application | WesternBlotting |
BackgroundMutationsinelongationofverylong-chainfattyacid-4(ELOVL4)areassociatedwithautosomaldominantStargardt-likemaculardegeneration(STGD3),withafivebase-pair(5bp)deletionmutationresultinginthelossof51carboxy-terminalaminoacidsandtruncationoftheprotein.Elovl4isexpressedintheretinaandonlyalimitednumberofmammalianothertissues,includingskin,withunknownfunction.Inamousemodelwiththe5-bpdeletionintheElovl4gene,miceintheheterozygousstate(Elovl4(+/del))demonstrateprogressivephotoreceptordegeneration.Homozygousmice(Elovl4(del/del))displayscaly,wrinkledskin,haveseverelycompromisedepidermalpermeABIlitybarrierfunction,anddiewithinafewhoursafterbirth.LipidanalysesofepidermisfromElovl4(del/del)miceshowadecreaseinverylong-chainfattyacids(VLFAs)inboththeceramide/glucosylceramideandthefreefatty-acidfractions.ELOVL4isrequiredforgeneratingVLFAcriticalforepidermalbarrierfunction.
Synonyms:Elongationofverylongchainfattyacidsprotein4,ELOV4
Source
Immunogen:SyntheticpeptdidederivedfromthehumanELOV4protein
Product
ProductForm:Unconjugated
Formulation:Providedassolutioninphosphatebufferedsalinewith0.08%sodiumazide
Concentration:Seevialforconcentration
ApplicationsForusewithELOV4polyclonalantibodies(Cat.No.X2087P&X2381P).
FunctionalAnalysis:WesternBlotting
PositiveControl:Expressedintheretinaandatmuchlowerlevelinthebrain.
StorageProductshouldbestoredat-20ºC.Aliquottoavoidfreeze/thawcycles
ProductStability:Productsarestableforoneyearfrompurchasewhenstoredproperly
ShippingConditions:Shipongelice,storeat-20ºCimmediatelyuponarrival
CautionThisproductisintendedFORRESEARCHUSEONLY,andFORTESTSINVITRO,notforuseindiagnosticortherapeuticproceduresinvolvinghumansoranimals.Itmaycontainhazardousingredients. PleaserefertotheSafetyDataSheets(SDS)foradditionalinformationandproperhandlingprocedures.Disposeproductremaindersaccordingtolocalregulations.Thisdatasheetisasaccurateasreasonablyachievable,butNordic-Mubioacceptsnoliabilityforanyinaccuraciesoromissionsinthisinformation.
References1:KobayashiT,etal.ELOVL2overexpressionenhancestriacylglycerolsynthesisin3T3-L1andF442Acells.FEBSLett.2007Jul10;581(17):3157-63.Epub2007Jun11.2:DeAngelisMM,etal.Cigarettesmoking,CFH,APOE,ELOVL4,andriskofneovascularage-relatedmaculardegeneration.ArchOphthalmol.2007Jan;125(1):49-54.3:VasireddyV,etal.LossoffunctionalELOVL4depletesverylong-chainfattyacids(>or=C28)andtheuniqueomega-O-acylceramidesinskinleADIngtoneonataldeath.HumMolGenet.2007Mar1;16(5):471-82.Epub2007Jan5.4:SridharanS,etal.X-raycrystalstructureofMycobacteriumtuberculosisbeta-ketoacylacylcarrierproteinsynthaseII(mtKasB).JMolBiol.2007Feb16;366(2):469-80.Epub2006Nov7.5:SeitsonenS,etal.AnalysisofvariantsinthecomplementfactorH,theelongationofverylongchainfattyacids-like4andthehemicentin1genesofage-relatedmaculardegenerationintheFinnishpopulation.MolVis.2006Jul20;12:796-801.6:JakobssonA,etal.Fattyacidelongasesinmammals:theirregulationandrolesinmetabolism.ProgLipidRes.2006May;45(3):237-49.Epub2006Mar6.Review.7:PleiteR,etal.Inhibitorsoffattyacidbiosynthesisinsunflowerseeds.JPlantPhysiol.2006Sep;163(9):885-94.Epub2006Feb28.8:KajikawaM,etal.Isolationandfunctionalcharacterizationoffattyaciddelta5-elongasegenefromtheliverwortMarchantiapolymorphaL.FEBSLett.2006Jan9;580(1):149-54.Epub2005Dec6.9:KooAJ,etal.Identificationofaplastidacyl-acylcarrierproteinsynthetaseinArabidopsisanditsroleintheactivationandelongationofexogenousfattyacids.PlantJ.2005Nov;44(4):620-32.10:ZhangK.,etal.A5-bpdeletioninELOVL4isassociatedwithtworelatedformsofautosomaldominantmaculardystrophy.;Nat.Genet.27:89-93(2001).11:EdwardsA.O.,etal;AnovelgeneforautosomaldominantStargardt-likemaculardystrophywithhomologytotheSUR4proteinfamily.;Invest.Ophthalmol.Vis.Sci.42:2652-2663(2001).
ProteinReference(s)
DatabaseName:UniProt
Accessionnumber:Q9GZR5(Human)
SpeciesAccession:Human
SafetyDatasheet(s)forthisproduct:NM_SodiumAzide/wp-content/uploads/SDS/AntibodySDSwithSodiumAzideNoridic-MUbio.pdf
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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.

