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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Cell Meter™ TUNEL Apoptosis Assay Kit *Red Fluorescence*/22844/50 Tests
产品编号:22844
市  场 价:¥85560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$4278.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Cell Meter™ TUNEL Apoptosis Assay Kit *Red Fluorescence*/22844/50 Tests
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)556/579
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
DNAfragmentationrepresentsacharacteristicoflatestageapoptosis.DNAfragmentationinapoptoticcellscanbedetectedbyterminaldeoxynucleotidyltransferase(TdT)-mediateddUTPnickendlabeling(TUNEL).TheTUNELassayreliesonthepresenceofnicksintheDNAwhichcanbeidentifiedbyTdT,anenzymethatcatalyzestheadditionofdUTPsthataresecondarilylabeledwithaMarker.AlltheexistingTUNELassayscontainthehighlytoxicsodiumcacodylatewhichmightinducesapoptosisandalsodecreaseDNAproductionandDNAstrands.OurCellMeter™TUNELApoptosisAssayKitusesproprietarybuffersystemfreeofsodiumcacodylate.ThekitisbasedontheincorporationofouruniqueproprietaryfluorescentdyeintotheDNAfragmentsthatformduringapoptosis.Theassayisoptimizedforthedirectdetectionofapoptosisineitherdetachedorattachedcellswithoutusingantibody.Thekitprovidesalltheessentialcomponentswithanoptimizedassayprotocol.Itissuitableforfluorescencemicroplatereader,fluorescencemicroscope,orflowcytometer.ItssignalcanbeeasilydetectedatEx/Em=550nm/590nm.
SpectrumAdvancedSpectrumViewer

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Movemouseovergridtodisplaywavelength&intensityvalues.

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

Note:ThawComponentsCatroomtemperature,keepComponentsAandBonicebeforeuse.

 

1.      Culturecellstoanoptimaldensityforapoptosisinductionaccordingtoyourspecificprotocol.Werecommendabout30,000to50,000cells/wellforadherentcellsgrownina96-wellmicroplateculture,orabout1to2x106cells/mLfornon-adherentcells. Atthesametime,cultureanon-inducednegativecontrolcellpopulationatthesamedensityastheinducedpopulationforeverylabelingcondition. 
Note:WetreatedHeLacellswith100nM-1µMstaurosporinefor4hourstoinducecellapoptosis.SeeFigure1fordetails.

2.      FixationandPermeABIlization

a   Removecellmedia.

b   Add100μL/well/96-wellplateof4%formaldehydefixativebuffer(notsupplied)toeachwell.

Note:Fornon-adherentcells,adddesiredamount(suchas2x106cells/mL)of4%formaldehydefixativebuffer.

c   Incubateplatesfor20to30minutesatroomtemperature.

d   Removefixative.

Optional:add100μL/well/96-wellplateofthepermeabilizationreagent(0.2%TritonX-100inPBS,notsupplied)afterthefixationifneeded,andincubatetheplatefor10minutesatroomtemperature.

e   WashthecellswithPBS2-3times.

Optional:YoumayalsoprepareapositivecontrolforTUNELreactionusingDNAaseIbydigestingcellswithDNAaseIfor30minatroomtemperaturebeforeproceedtoTUNELreaction(Step3)

 

3.      TUNELreaction

3.1   Preparereactionmixturejustbeforeusebasedonthenumberofsamplestobeassayed:

 

ReactionComponents

Volume PerWell

100XTunnelyte™Red(ComponentA)

0.5µL

ReactionBuffer(ComponentB)

50µL

Totalvolume

50.5µL

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.

3.2   Add50µLofthereactionmixture(fromStep3.1)toeachwellortubeandincubateat37°Cfor60minutes.

3.3   Removethereactionmixture,andwashthecells3-5timeswith200µL/wellofPBS.

 

4.      Monitorthefluorescenceintensitybyfluorescencemicroscope,flowcytometer,orfluorescencemicroplatereaderatEx/Em=550/590-650nm.

5.      Optional:Stainthenucleuswith1XHoechst(ComponentC,Ex/Em=350/460nm)forimageanalysis.

References&Citations
CitationExplorer

Vaccarinalleviateshypertensionandnephropathyinrenovascularhypertensiverats
Authors:WeiweiCai,ZhenpengZhang,YiqiHuang,HaijianSun,LiyingQiu
Journal:ExperimentalandTherapeuticMedicine(2018):924--932

CO-releasingmolecules-2attenuatesox-LDL-inducedinjuryinHUVECsbyamelioratingmitochondrialfunctionandinhibitingWnt/β-cateninpathway
Authors:Hai-JianSun,Dong-YanXu,Yi-XinSun,TongXue,Chen-XingZhang,Zhi-XuanZhang,WeiLin,Ke-XueLi
Journal:BiochemicalandBiophysicalResearchCommunications(2017)

Salusin-βmediateshighglucose-inducedendothelialinjuryviadisruptionofAMPKsignalingpathway
Authors:XuexueZhu,YuetaoZhou,WeiweiCai,HaijianSun,LiyingQiu
Journal:BiochemicalandBiophysicalResearchCommunications(2017)

VaccarinprotectshumanmicrovascularendothelialcellsfromapoptosisviaattenuationofHDAC1andoxidativestress
Authors:XuexueZhu,YueyueLei,FanggenTan,LeileiGong,HaifengGong,WeiYang,TingChen,ZhixuanZhang,WeiweiCai,BaoHou
Journal:EuropeanJournalofPharmacology(2017)

AxlisrequiredforTGF-β2-induceddormancyofprostatecancercellsinthebonemarrow
Authors:KenjiYumoto,MatthewREber,JingchengWang,FrankCCackowski,AnnMDecker,EunsohlLee,AnaRitaNobre,JulioAAguirre-Ghiso,YounghunJung,RussellSTaichman
Journal:ScientificReports(2016)

GrowthArrest-Specific6(GAS6)PromotesProstateCancerSurvivalbyG1Arrest/SPhaseDelayandInhibitionofApoptoticPathwayDuringChemotherapyinBoneMarrow
Authors:EunsohlLee,AnnMDecker,FrankCCackowski,LuliaAKana,KenjiYumoto,YounghunJung,JingchengWang,LauraButtitta,ToddMMorgan,RussellSTaichman
Journal:Journalofcellularbiochemistry(2016)

RFX1--dependentactivationofSHP-1inducesautophagybyanovelobatoclaxderivativeinhepatocellularcarcinomacells
Authors:Jung-ChenSu,Ping-HuiTseng,Cheng-YiHsu,Wei-TienTai,Jui-WenHuang,Ching-HuaiKo,Mai-WeiLin,Chun-YuLiu,Kuen-FengChen,Chung-WaiShiau
Journal:Oncotarget(2014):4909


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