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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/Hydroethidine [Dihydroethidium] *CAS 104821-25-2*/15202/5X1 mg
产品编号:15202
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AAT Bioquest/Hydroethidine [Dihydroethidium] *CAS 104821-25-2*/15202/5X1 mg
商品介绍
Overview
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Ex/Em(nm)518/605
MW315.41
CAS#104821-25-2
SolventDMSO
StorageF/D/L
CategoryEnzymeDetection
RedoxEnzymes
RelatedCellSignalingMolecules
ReactiveOxygenSpecies
Hydroethidineoperateseffectivelyasaprobeformeasurementofreactiveoxygenspecies.Thedyeenterscellsfreelyandisdehydrogenatedtoanethidiumcompound.TheprobehasbeenusedextensivelywithNKcellandasavitaldyeforidentificationofproliferationandhypoxiccellsintumors.StudieshavebeenperformedusingneutrophilsandendothelialcellsaswellasHL60cellsandmacrophages.AmajoradvantageofthisprobeisitsABIlitytodistinguishbetweensuperoxideandH2O2.Fluorescenceemissionoccursataround600nm.
SpectrumAdvancedSpectrumViewer

Sorry,yourbrowserdoesnotsupportinlineSVG.RelativeIntensity(%)100806040200Sorry,yourbrowserdoesnotsupportinlineSVG.
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Movemouseovergridtodisplaywavelength&intensityvalues.

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Make5-10mMDMSOstocksolution.TheunusedDMSOstocksolutionshouldbealiquotedintoasingleusevialandstoredat-20°C.Keepfromlight.
  2. Makethedyeworkingconcentrationof5-20µMinaphysiologicalbuffer(suchasPBS,HBSS,HEPES).Theoptimalworkingconcentrationforyourapplicationmustbeempiricallydetermined.
  3. Addequalvolume(suchas100µLofthecellsingrowthmedium)ofthedyeworkingsolution(fromStep2)tothecells,andincubatethecellsatRTor37oCfor5to60minutes.
  4. Determinethebaselinefluorescenceintensityofasampleoftheloadedcellspriortoexposingthecellstoexperimentalinducements.
  5. Negativecontrolsshouldbeassessedasfollows:

5.1    Examinethefluorescenceofcell-freemixturesofdyeandbuffer/mediawithandwithouttheinducer.Intheabsenceofextracellularesterasesandotheroxidativeenzymes,thegradualincreaseinfluorescenceovertimemayberelatedtospontaneoushydrolysis,atmosphericoxidation,and/orlight-inducedoxidation.

5.2    Examinethefluorescenceofuntreated(control)loadedcellsthathavebeenmaintainedingrowthmediumorbuffer.Inhealthycells,oxygenrADIcalsareeliminatedbycellularenzymesand/ornaturalantioxidants.Followingthedye-loadingrecoveryperiod,healthycellsshouldexhibitalowleveloffluorescencethatisrelativelystableforthedurationoftheexperiment;however,agradualincrease(duetoauto-oxidation)ordecrease(duetolossofdyefromcellsorphotobleaching)influorescencemaybeobserved.Intheabsenceofanystimulusorinducement,aburstoffluorescenceinhealthy,untreatedcellscouldindicateprogresstocelldeathorsomeotheroxidativeevent.

  1. Positivecontrolsmaybestimulatedwithtert-butylhydroperoxide(TBHP)toafinalconcentrationof~100µM(increaseordecreasedosebasedonthesensitivityandresponseofthecells).
References&Citations
CitationExplorer

RoleofepigeneticregulationontheinductionofapoptosisinJurkatleukemiacellsbywhitegrapepomacerichinphenoliccompounds
Authors:AntonioJLeón-González,MJoséJara-Palacios,MalakAbbas,FJHeredia,ValérieSchini-Kerth
Journal:Food&Function(2017)

AstragalosideIVimprovestheisoproterenol-inducedvasculardysfunctionviaattenuatingeNOSuncoupling-mediatedoxidativestressandinhibitingROS-NF-$kappa$Bpathways
Authors:ChonghuaXu,FutianTang,MeiliLu,JingYang,RonghuiHan,MengMei,JinHu,MingshengZhou,HongxinWang
Journal:Internationalimmunopharmacology(2016):119--127

InhibitoryeffectofSophorasubprosratepolysaccharideonmitochondriaoxidativestressinducedbyPCV-2infectioninRAW264.7cells
Authors:Zi-JieSu,JianYang,Wen-JuanLuo,Ying-YiWei,Xue-HongShuai,Ting-JunHu
Journal:InternationalJournalofBIOLOGicalMacromolecules(2016)

EvaluationoftheMedicinalHerbGraptopetalumparaguayenseasaTreatmentforLiverCancer
Authors:Wei-HsiangHsu,Chia-ChuanChang,Kai-WenHuang,Yi-ChenChen,Shih-LanHsu,Li-ChenWu,Ann-PingTsou,Jin-MeiLai,Chi-YingFHuang
Journal:PloSone(2015):e0121298

SalvagingbrainischemiabyincreasingneuroprotectantuptakeviananoagoNISTmediatedbloodbrainbarrierpermeabilityenhancement
Authors:ShuyanZheng,Ying-YingBai,YikangLiu,XihuiGao,YanLi,YinzhiChangyi,YuanchengWang,DiChang,ShenghongJu,CongLi
Journal:Biomaterials(2015):9--20

PretreatmentwithAstragalosideIVprotectshumanumbilicalveinendothelialcellsfromhydrogenperoxideinducedoxidativestressandcelldysfunctionviainhibitingeNOSuncouplingandNADPHoxidase-ROS-NF-$kappa$Bpathway
Authors:ChonghuaXu,FutianTang,MeiliLu,JingYang,RonghuiHan,MengMei,JinHu,HongxinWang
Journal:CanadianJournalofPhysiologyandPharmacology


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