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AAT Bioquest/Hydroethidine [Dihydroethidium] *CAS 104821-25-2*/15202/5X1 mg
Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 518/605 |
MW | 315.41 |
CAS# | 104821-25-2 |
Solvent | DMSO |
Storage | F/D/L |
Category | EnzymeDetection RedoxEnzymes |
Related | CellSignalingMolecules ReactiveOxygenSpecies |
Spectrum | AdvancedSpectrumViewer |
- Make5-10mMDMSOstocksolution.TheunusedDMSOstocksolutionshouldbealiquotedintoasingleusevialandstoredat-20°C.Keepfromlight.
- Makethedyeworkingconcentrationof5-20µMinaphysiologicalbuffer(suchasPBS,HBSS,HEPES).Theoptimalworkingconcentrationforyourapplicationmustbeempiricallydetermined.
- Addequalvolume(suchas100µLofthecellsingrowthmedium)ofthedyeworkingsolution(fromStep2)tothecells,andincubatethecellsatRTor37oCfor5to60minutes.
- Determinethebaselinefluorescenceintensityofasampleoftheloadedcellspriortoexposingthecellstoexperimentalinducements.
- 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.
- Positivecontrolsmaybestimulatedwithtert-butylhydroperoxide(TBHP)toafinalconcentrationof~100µM(increaseordecreasedosebasedonthesensitivityandresponseofthecells).
References&Citations | CitationExplorer |
RoleofepigeneticregulationontheinductionofapoptosisinJurkatleukemiacellsbywhitegrapepomacerichinphenoliccompounds
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Journal:Internationalimmunopharmacology(2016):119--127
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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