Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 490/530 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | EnzymeDetection HorserADIshPeroxidase(HRP) |
Related | ReactiveOxygenSpecies MicroplateReaders BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1.Preparecells:
Foreachsample,preparecellsin0.5mLgrowthmediumorbufferofyourchoiceatadensityof5×105to1×106cells/mL.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforhydrogenperoxideinduction.
2.PrepareOxiVision™Greenperoxidesensorstocksolution:
Add100µLofDMSO(ComponentB)intothevialofOxiVision™Greenperoxidesensor(ComponentA),andmixthemwell.
Note:1µLofreconstitutedOxiVision™Greenperoxidesensorstocksolutionisfor0.5mLcells.Thestocksolutionshouldbeusedpromptly.Anyremainingsolutionshouldbealiquotedandfrozenat-20oC.Avoidrepeatedfreeze-thawcyclesandprotectfromlight.
3.Runthehydrogenperoxideassay:
3.1 StaincellswithOxiVision™Greenperoxidesensorinfullmediumorinyourdesiredbufferat37°Cfor30minutes,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 Alternatively,treatcellswithtestedcompoundsat37°Cfordesiredperiodoftime(asinStep3.2).Removethetreatmentsolution,thenstaincellswithOxiVision™Greenperoxidesensorinfullmediumorinyourdesiredbufferat37°Cfordesiredperiodoftime.
3.4 MonitorthefluorescenceintensityatFITCchannel(Ex/Em=490/530nm)usingaflowcytometer.Gateonthecellsofinterest,excludingdebris.
References&Citations | CitationExplorer |
Plasma-activatedmediumselectivelyeliminatesundifferentiatedhumaninducedpluripotentstemcells
Authors:RyoMatsumoto,KazunoriShimizu,TakunoriNagashima,HiromasaTanaka,MasaakiMizuno,FumitakaKikkawa,MasaruHori,HiroyukiHonda
Journal:RegenerativeTherapy(2016):55--63