Host/IsotypeMouse/IgG1 Clone19C4B2.1 SpeciesReactivityAllspecies ValidationDataAc-KAntibodyWhitePaper | AAC03,anti-acetyllysineantibodyisapan-acetyllysinemousemonoclonalantibodythatispartoftheSignal-Seeker™productline.TheAnti-acetyl-lysineantibodyrecognizesproteinspost-translationallymodifiedbyacetylationontheepsilonaminegroupsoflysineresiduesthatoccuron30-50%ofallproteinsandinparticularhistones,p53,tubulinandmyosin.AproprietarymixtureofacetylatedproteinswasusedtoproduceahighlyrobustantibodythathasbeenshowntorecognizeawiderangeofacetylatedproteinsinIP,WB,ChIPandIFapplications.ThisAnti-acetyl-lysineantibodyhasmanyadvantageswhencomparedtoothercommerciallyavailableantibodiesasshownbelow. |
ValidatedApplications
WesternBlotusingAcetyl-LysineAntibody(AAC03) Fig1:A:Murinetissueextract,30mgbrainextract.B:30mgofCos-7celllysatetreatedwithTSAandnicotinamide(+)oruntreated(-).Stronglyenhancedbandsat55and14-16kDainTSA-treatedlysatecorrespondtoacetylatedtubulinandhistoneproteins,respectively.C:TitrationofacetylatedBSA.Lanes1-5contain0.5,0.1,0.05,0.01,and0.005ngAc-BSA,lanes6-7contain500and1000ngnon-acetylatedBSA,respectively.AAC03wasusedata1:500dilutionfollowingtherecommendedwesternblotprotocol. ToseethefullWesternblotcomparison,seetheOptimizedProtocolsortheproductdatasheet. | |
ImmunoprecipitationusingAcetyl-LysineAntibody Fig2:Cos-7cellswereeithertreated(+)oruntreated(-)withTSA(1mM)andnicotinamide(1mM)for6hours.CelllysateswerepreparedinBlastRbufferandfiltersystemand1mgoflysateperreactionwasusedforIPofacetylatedproteins.20mlofAAC03wasusedperIPreaction.WesternblotsofimmunoprecipitatedproteinsweredevelopedusingAAC03-HRPat1:3000dilution. ToseethefullImmunoprecipitationcomparison,seetheOptimizedProtocolsortheproductdatasheet. | |
ImmunofluorescenceusingAcetyl-LysineAntibody Fig3:Swiss3T3cells,untreated(aandc)ortreated(bandd)withTSA(1mMfor6h),werestainedasdescribedinthedatasheet.Acetylatedproteinswerevisualizedusingagreenfluorescentsecondary.ActinfiberswerevisualizedusingaredRhodaminePhalloidinandthenucleuswasstainedwithDAPI.TheacetylatedmicrotubulenetworkisclearlyvisIBLewithTSA-treatment,whilethegreenfluorescentnuclearintensityindicatethehighabundanceofacetylatedproteinsinthenucleus.Incandd,acetylatedBSA(10mg/ml)wasusedtocompeteforAAC03bindingasanindicatorofAAC02specificityforacetyl-lysinemodifications. ToseethefullImmunofluorescenceprotocol,seetheOptimizedProtocolsortheproductdatasheet. | |
ChIPusingAcetyl-LysineAntibody Fig4:UtilizationofAAC03forChIP.ChromatinwaspreparedfromA431cells,eitheruntreatedortreatedwithTSA(1mM)andnicotinamide(1mM)for6hours.ChIPwasperformedasdescribed.mIgG:mouseIgGusedforChIPcontrol;AAC02:anti-acetyllysineantibodyusedforChIP;Input:celllysatepriortoChIP;H2O:WaterusedasPCRcontrol.ThePCRproductsobtainedwithGAPDHprimersare166bp. ToseetherecommendedChIPprotocol,seetheOptimizedProtocolsortheproductdatasheet. |
ProteinAcetylationBackground
Acetylationofproteinscanoccurasaco-translationalorpost-translationalmodification(PTM)(1).Co-translationalacetylationoccursattheN-terminalofapproximately85%ofmammalianproteins,itisirreversibleandisthoughttobeimportantinproteinstABIlity,localizationandsynthesis(1).Post-translationalacetylationoccursontheepsilonaminogroupoflysineresiduesasareversibleandhighlydynamicPTMthatisknowntobeakeyregulatorinmultiplecellularevents,includingchromatinstructure,transcription,metabolism,signaltransductionandcytoskeletalregulation(2-3).Todateover4,000proteinshavebeenidentifiedastargetsforPTMacetylationwhichiscomparabletophosphorylationincellularprevelance(3).AntibodyAAC01detectsacetyllysinePTMs.
References
1BogdanP.andShermanF.2002.Thediversityofacetylatedproteins.GenomeBiol.3(5):reviews0006.
2LundbyA.etal.2012.Proteomicanalysisoflysineacetylationsitesinrattissuesrevealsorganspecificityandcellularpatterns.CellReports2:419-431.
3SadoulK.etal.2010.Thetaleofproteinlysineacetylationinthecytoplasm.J.Biomed.Biotech.2011:1-15.
4GolemisEAet.Al,Protein-ProteinInteractions,CSHLP,2005,p67
Formoreinformationcontact:signalseeker@Cytoskeleton.com
AssociatedProducts:
Signal-Seeker™Acetyl-LysineDetectionKit(Cat.#BK163)
Signal-Seeker™:BlastR™RapidLysatePrepKit(Cat.#BLR01)
Signal-Seeker™AcetylationAffinityBeads(Cat.#AAC04-beads)
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以下来自摆渡百科
细胞在发生凋亡时,会激活一些DNA内切酶,这些内切酶会切断核小体间的基因组DNA。细胞凋亡时抽提DNA进行电泳检测,可以发现180-200bp的DNA ladder。基因组DNA断裂时,暴露的3’-OH可以在末端脱氧核苷酸转移酶(Terminal
Deoxynucleotidyl Transferase, TdT) 的催化下加上荧光素 (FITC) 标记的dUTP
(fluorescein-dUTP) ,从而可以通过荧光显微镜或流式细胞仪进行检测,这就是TUNEL (TdT-mediated dUTP
Nick-End Labeling) 法检测细胞凋亡的原理。
想买消炎药怎么办?
去药店买药想买不是抗生素的消炎药,那工作人员说不是抗生素怎么消炎呢,请问知情网友求一替代品,谢了。
sc-153是兔抗大鼠ERK2的多克隆抗体
还有很多关于ERK1和ERK2的抗体
若想知道更多信息,你可以拨打Santa cruz 上海分公司的电话咨询
021-6093-6351
我们会按照你的实验需求推荐最适合你的产品。
第二个问题:兔子不能免疫兔子的。免疫的一个重要概念是识别“自己”和“非己”,如果对自己的蛋白产生免疫反应,那就麻烦了。
可以类比器官移植,亲缘关系越近,越不容易产生免疫排斥。
你看这个抗体的质量怎么样,说明里面有没有说可以做组化,还是只能做western blot。