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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*/22837/100 Tests
商品详细AAT Bioquest/Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*/22837/100 Tests
AAT Bioquest/Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*/22837/100 Tests
AAT Bioquest/Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*/22837/100 Tests
商品编号: 22837
品牌: aatbio
市场价: ¥56560.00
美元价: 33936.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)651/662
MWN/A
CAS#N/A
SolventN/A
StorageR/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
BiochemicalAssays
AnnexinVmaybeconjugatedtoFluorochromesincludingAPC.Thisformatretainsitshighaffinityforphosphatidylserine(PS)andthusservesasasensitiveprobeforflowcytometricanalysisofcellsthatareundergoingapoptosis.SinceexternalizationofPSoccursintheearlierstagesofapoptosis,APCAnnexinVstainingcanidentifyapoptosisatanearlierstagethanassaysbasedonnuclearchangessuchasDNAfragmentation.APCAnnexinVstainingprecedesthelossofmembraneintegritywhichaccompaniesthelateststagesofcelldeathresultingfromeitherapoptoticornecroticprocesses.Therefore,stainingwithAPCAnnexinVistypicallyusedinconjunctionwithavitaldyesuchaspropidiumiodide(PI)or7-Amino-Actinomycin(7-AAD)toallowtheinvestigatortoidentifyearlyapoptoticcells(7-AADnegative,APCAnnexinVpositive).Viablecellswithintactmembranesexclude7-AAD,whereasthemembranesofdeadanddamagedcellsarepermeableto7-AAD.Forexample,cellsthatareconsideredviablearebothAPCAnnexinVand7-AADnegativewhilecellsthatareinearlyapoptosisareAPCAnnexinVpositiveand7-AADnegative,whilecellsthatareinlateapoptosisoralreadydeadarebothAPCAnnexinVand7-AADpositive.Thisassaydoesnotdistinguishbetweencellsthathaveundergoneapoptoticdeathversusthosethathavediedasaresultofanecroticpathwaybecauseineithercase,thedeadcellswillstainwithbothAPCAnnexinVand7-AAD.However,whenapoptosisismeasuredovertime,cellscanbeoftentrackedfromAPCAnnexinVand7-AADnegative(viable,ornomeasurableapoptosis),toAPCAnnexinVpositiveand7-AADnegative(earlyapoptosis,membraneintegrityispresent)andfinallytoAPCAnnexinVand7-AADpositive(endstageapoptosisanddeath).Themovementofcellsthroughthesethreestagessuggestsapoptosis.Incontrast,asingleobservationindicatingthatcellsarebothAPCAnnexinVand7-AADpositive,inofitself,revealslessinformationabouttheprocessbywhichthecellsunderwenttheirdemise.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.      Treatcellswithtestcompoundsforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithcamptothecin)toinduceapoptosis.

Note:AnnexinVflowcytometricanalysisonadherentcellsisnotroutinelytestedsincespecificmembranedamagemayoccurduringcelldetachmentorharvesting.However,methodsforutilizingAnnexinVforflowcytometryonadherentcelltypeshavebeenpreviouslyreportedbyCasiola-Rosenetal.andvanEngelendetal(seeRefs1and2).

2.      Centrifugethecellstoget1-5×105cells/tube.

3.      ResUSPendcellsin200μLofAssayBuffer(ComponentB).

4.      ResuspendtheAPC-AnnexinV(ComponentA)with200μLPBSwith0.2%BSAtohave100Xstocksolution.

Note:Storethereconstitutedstocksolutionat4oC.DoNotFreeze.

5.      Add2μLof100XAPC-AnnexinVstocksolution(fromStep4)intothecells.

Optional:Add2µLof100XPropidiumIodide(ComponentC)fornecrosiscells.

6.      Incubateatroomtemperaturefor20to60minutes,protectedfromlight.

7.      Add200to300μLofAssayBuffer(ComponentB)toincreasevolumebeforeanalyzingthecellswithaflowcytometer(SeeStep7).

8.      MonitorthefluorescenceintensityofAPC-AnnexinVusingtheFL4channel(Ex/Em=635/660nm),andmeasurethecellviABIlitywithpropidiumiodideusingtheFL2channel.

References&Citations
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1.   BaeON,LimKM,NohJY,ChungSM,KimSH,ChungJH.(2009)Trivalentmethylatedarsenical-inducedphosphatidylserineexposureandapoptosisinplateletsmayleadtoincreasedthrombusformation.ToxicolApplPharmacol,239,144.

2.   BurteaC,LaurentS,LancelotE,BalletS,MurariuO,RousseauxO,PortM,VanderElstL,CorotC,MullerRN.(2009)PeptidictargetingofphosphatidylserinefortheMRIdetectionofapoptosisinatheroscleroticplaques.MolPharm,6,1903.

3.   DongHP,HolthA,KleinbergL,RuudMG,ElstrandMB,TropeCG,DavidsonB,RisbergB.(2009)Evaluationofcellsurfaceexpressionofphosphatidylserineinovariancarcinomaeffusionsusingtheannexin-V/7-AADassay:clinicalrelevanceandcomparisonwithotherapoptosisparameters.AmJClinPathol,132,756.

4.   LiuT,ZhuW,YangX,ChenL,YangR,HuaZ,LiG.(2009)DetectionofapoptosisbasedontheinteractionbetweenannexinVandphosphatidylserine.AnalChem,81,2410.

5.   MirnikjooB,BalasubramanianK,SchroitAJ.(2009)Suicidalmembranerepairregulatesphosphatidylserineexternalizationduringapoptosis.JBiolChem,284,22512.

6.   MirnikjooB,BalasubramanianK,SchroitAJ.(2009)Mobilizationoflysosomalcalciumregulatestheexternalizationofphosphatidylserineduringapoptosis.JBiolChem,284,6918.

7.   ThapaN,KimS,SoIS,LeeBH,KwonIC,ChoiK,KimIS.(2008)Discoveryofaphosphatidylserine-recognizingpeptideanditsutilityinmolecularimagingoftumourapoptosis.JCellMolMed,12,1649.

8.   AiresV,HichamiA,FilomenkoR,PleA,RebeC,BettaiebA,KhanNA.(2007)Docosahexaenoicacidinducesincreasesin[Ca2+]iviainositol1,4,5-triphosphateproductionandactivatesproteinkinaseCgammaand-deltaviaphosphatidylserinebindingsite:implicationinapoptosisinU937cells.MolPharmacol,72,1545.

9.   BalasubramanianK,MirnikjooB,SchroitAJ.(2007)Regulatedexternalizationofphosphatidylserineatthecellsurface:implicationsforapoptosis.JBiolChem,282,18357.

10.   CauchonN,LangloisR,RousseauJA,TessierG,CadoretteJ,LecomteR,HuntingDJ,PavanRA,ZeislerSK,vanLierJE.(2007)PETimagingofapoptosiswith(64)Cu-labeledstreptavidinfollowingpretargetingofphosphatidylserinewithbiotinylatedannexin-V.EurJNuclMedMolImaging,34,247.


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品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
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淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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