Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 490/514 |
MW | ~600 |
CAS# | N/A |
Solvent | DMSO |
Storage | F/D/L |
Category | NeuroBIOLOGy ReactiveOxygenSpecies |
Related | CellFunctionalAnalysis RedoxEnzymes BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1.PrepareOxiVisionGreenTMhydrogenperoxidesensorworkingsolution:
1.1 Preparea2to5mMstocksolutionofOxiVisionGreenTMhydrogenperoxidesensorinhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionshouldbealiquotedandfrozenat-20oC.
Note:Avoidrepeatedfreeze-thawcycles.
1.2 Preparea2XOxiVisionGreenTMhydrogenperoxidesensorworkingsolution:Onthedayoftheexperiment,eitherdissolveOxiVisionGreenTMhydrogenperoxidesensorsolidinDMSOorthawanaliquotofthesensorstocksolutiontoroomtemperature.Preparea2Xworkingsolutionattheconcentrationrangingfrom2to20µMin20mMHepesbufferorbufferofyourchoice,pH7.ItisrecommendedtouseOxiVisionGreenTMhydrogenperoxidesensoratthefinalconcentrationof5µMtomeasureH2O2concentrationinsolution.
2.RunH2O2Assayinsupernatants:
2.1 Add50µLof2XOxiVisionGreenTMhydrogenperoxidesensorworkingsolution(fromStep1.2)toeachwelloftheH2O2standard,blankcontrol,andtestsamplestomakethetotalH2O2assayvolumeof100 µL/well.
Note:Fora384-wellplate,add25µLofsampleand25µLof2XOxiVisionGreenTMhydrogenperoxidesensorworkingsolutionintoeachwell.
2.2 Incubatethereactionatroomtemperaturefor15to60minutes,protectedfromlight.
2.3 MonitorthefluorescenceincreasewithafluorescenceplatereaderatEx/Em=490/525nm.
2.4 Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththeH2O2reactions.
3.RunH2O2AssayinLiveCells:
OxiVisionGreenTMhydrogenperoxidesensorcanbeloadedpassivelyintolivingcellsandreportthemicromolarchangesinintracellularH2O2concentrations.Thefollowingisasuggestedmicroscopeimagingprotocolwhichcanbemodifiedaccordingtoyourspecificresearchneeds.
3.1 TheOxiVisionGreenTMhydrogenperoxidesensorworkingsolutionshouldbepreparedasStep1.2.ItisrecommendedtousePBSorHanksBalancedSaltSolution(HBSS)with20mMHepesbufferinsteadof20mMHepesbufferonly.
3.2 Treatthecellsasdesired.
3.3 IncubatethecellswithOxiVisionGreenTMhydrogenperoxidesensorworkingsolutionfor5to60minoradesiredperiodoftime.WashthecellswithPBSbuffertwice.
3.4 MonitorthefluorescenceincreaseatEx/Em=490/525nmwithafluorescenceplatereaderwithbottomreadmode.OrimagethefluorescencechangebyafluorescencemicroscopyusingtheFITCchannel.
References&Citations | ![]() CitationExplorer |
Semaphorin4Dinhibitsneutrophilactivationandisinvolvedinthepathogenesisofneutrophil-mediatedautoimmunevasculitis
Authors:MasayukiNishide,SatoshiNojima,DaisukeIto,HyotaTakamatsu,ShoheiKoyama,SujinKang,TetsuyaKimura,KeikoMorimoto,TakashiHosokawa,YoshitomoHayama
Journal:AnnalsoftheRheumaticDiseases(2017):annrheumdis--2016
Aggravationofbraininfarctionthroughanincreaseinacroleinproductionandadecreaseinglutathionewithaging
Authors:TakeshiUemura,KentaWatanabe,MisakiIshibashi,RyotaroSaiki,KyoshiroKuni,KazuhiroNishimura,ToshihikoToida,KeikoKashiwagi,KazueiIgarashi
Journal:Biochemicalandbiophysicalresearchcommunications(2016):630--635
Hydrogenperoxidedetectionwithhighspecificityinlivingcellsandinflamedtissues
Authors:LeiRong,ChiZhang,QiLei,Ming-MingHu,JunFeng,Hong-BingShu,YiLiu,Xian-ZhengZhang
Journal:RegenerativeBiomaterials(2016):rbw022
Modificationoflignininsugarcanebagassebyamonocopperhydrogenperoxide-generatingoxidasefromThermobifidafusca
Authors:Cheng-YuChen,Cheng-ChengLee,Hung-ShuanChen,Chao-HsunYang,Shu-PingWang,Jyh-HorngWu,MenghsiaoMeng
Journal:ProcessBiochemistry(2016):1486--1495
Dopamine-mediatedoxidationofmethionine127inα-synucleincausescytotoxicityandoligomerizationofα-synuclein
Authors:KazuhiroNakaso,NaokoTajima,SatoruIto,MariTeraoka,AtsushiYamashita,YosukeHorikoshi,DaisukeKikuchi,ShinsukeMochida,KenjiNakashima,TatsuyaMatsura
Journal:PLoSOne(2013):e55068
Hydrogenperoxidestimulatestheepithelialsodiumchannelthroughaphosphatidylinositide3-kinase-dependentpathway
Authors:He-PingMa
Journal:JournalofBiologicalChemistry(2011):32444--32453
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