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AAT Bioquest/Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Blue Fluorescence Optimized for Flow
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 405/450 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | EnzymeDetection HorserADIshPeroxidase(HRP) |
| Related | ReactiveOxygenSpecies MicroplateReaders BiochemicalAssays |
1.Preparecells:
Foreachsample,preparecellsin0.5mLgrowthmediumorbufferofyourchoiceatadensityof5×105to1×106cells/mL.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforhydrogenperoxideinduction.
2.PrepareOxiVision™Blueperoxidesensorstocksolution:
Add100µLofDMSO(ComponentB)intothevialofOxiVision™Blueperoxidesensor(ComponentA),andmixthemwell.
Note:1µLofreconstitutedOxiVision™Blueperoxidesensorstocksolutionisfor0.5mLcells.Thestocksolutionshouldbeusedpromptly.Anyremainingsolutionshouldbealiquotedandrefrozenat-20oC.Avoidrepeatedfreeze-thawcyclesandprotectfromlight.
3.Runthehydrogenperoxideassay:
3.1 StaincellswithOxiVision™Blueperoxidesensorinfullmediumorinyourdesiredbufferat37°Cfor20-30minutes,protectedfromlight.
3.2 Treatcellswithtestcompoundsinfullmediumorinyourdesiredbufferat37°Cfordesiredperiodoftime.Forcontrolsamples(untreatedcells),addthecorrespondingamountofcompoundbuffer.
Note1:It"srecommendedtotreatcellsinfullmedium.However,iftestedcompoundsareserumsensitive,growthmediumandserumfactorscanbeaspiratedawaybeforetreatment.ResUSPendcellsin1XHank"ssaltsolutionand20mMHepesbuffer(HHBS)orthebufferofyourchoiceafteraspiration.Alternatively,cellscanbetreatedinserum-freemedia.
Note2:WetreatedJurkatcellswith100µMhydrogenperoxideinfullmediumat37°Cfor90minutestoinducehydrogenperoxide.SeeFigure1fordetails.
3.3 MonitorthefluorescenceintensityatPacificBluechannel(Ex/Em=405/450nm)usingaflowcytometer.Gateonthecellsofinterest,excludingdebris.
| References&Citations |  CitationExplorer |
Plasma-activatedmediumselectivelyeliminatesundifferentiatedhumaninducedpluripotentstemcells
Authors:RyoMatsumoto,KazunoriShimizu,TakunoriNagashima,HiromasaTanaka,MasaakiMizuno,FumitakaKikkawa,MasaruHori,HiroyukiHonda
Journal:RegenerativeTherapy(2016):55--63
