Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 658/675 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | NeuroBIOLOGy ReactiveOxygenSpecies |
Related | CellSignaling CellFunctionalAnalysis BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1.Preparecells:
1.1 Foradherentcells:Platecellsovernightingrowthmediumat10,000to40,000cells/well/90µLfora96-wellplateor2,500to10,000cells/well/20µLfora384-wellplate.
1.2 Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandsUSPendthecellpelletsinculturemediumat50,000-100,000cells/well/90µLfora96-wellpoly-Dlysineplateor10,000-25,000cells/well/20µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortoyourexperiment.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.
2.PrepareROSBrite™670AssaySolution:
2.1 PrepareROSBrite™670stocksolution(500X):Add40µLofDMSO(ComponentC)intothevialofROSBrite™670(ComponentA),andmixthemwell.
Note:20µLofreconstitutedROSBrite™670stocksolutionisenoughfor1plate.Unusedportioncanbealiquotedandstoredat<-20°Cformorethanonemonthifthetubesaresealedtightlyandkeptfromlight.Avoidrepeatedfreeze-thawcycles.
2.2 PrepareROSBrite™670assaysolution:Add20µLof500XDMSOreconstitutedROSBrite™670stocksolution(fromStep2.1)into10mLofAssayBuffer(ComponentB),andmixthemwell.Thisassaysolutionisstableforatleast2hoursatroomtemperature.
3.RunROSAssay:
3.1 Treatcellswith10μLof10Xtestcompounds(96-wellplate)or5μLof5Xtestcompounds(384-wellplate)inyourdesiredbuffer(suchasPBSorHHBS).Forcontrolwells(untreatedcells),addthecorrespondingamountofcompoundbuffer.
3.2 ToinduceROS,incubatethecellplateatroomtemperatureorina5%CO2,37°Cincubatorforadesiredperiodoftime(forexample:30minutestreatmentforHelacellswith100µMtert-butylhydroperoxide(TBHP)).
3.3 Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofROSBrite™670workingsolution(fromStep2.2)intothecellplate.
3.4 Incubatethecellsina5%CO2,37°Cincubatorfor30mintoonehour,andmonitorthefluorescenceincreaseatEx/Em=650/675nm(cutoff=665m)withbottomreadmode,ortakeimageswithTRITCfilterset.
References&Citations | ![]() CitationExplorer |
Anti-proliferationeffectofbluelight-emittingdiodesagainstantibiotic-resistantHelicobacterpylori
Authors:JianweiMa,TakahiroHiratsuka,TsuyoshiEtoh,JunkoAkada,HajimeFujishima,NorioShiraishi,YoshioYamaoka,MasafumiInomata
Journal:JournalofGastroenterologyandHepatology(2017)
NotoginsenosideR1attenuateshighglucose-inducedendothelialdamageinratretinalcapillaryendothelialcellsbymodulatingtheintracellularredoxstate
Authors:ChunlanFan,YuanQiao,MinkeTang
Journal:DrugDesign,DevelopmentandTherapy(2017):3343
Goodhydrationandcell-biologicalperformancesofsuperparamagneticcalciumphosphatecementwithconcentration-dependentosteogenesisandangiogenesisinducedbyferriciron
Authors:JZhang,HSShi,JQLiu,TYu,ZHShen,JDYe
Journal:JournalofMaterialsChemistryB(2015):8782--8795
TopiramateProtectsPericytesfromGlucotoxicity:RoleforMitochondrialCAVAinCerebromicrovascularDiseaseinDiabetes
Authors:PingPatrick,TulinOPrice,AnaLDiogo,NaderSheibani,WilliamABanks,GulNShah
Journal:Journalofendocrinologyanddiabetes(2015)
Down-regulatedperoxisomeproliferator-activatedreceptorγ(PPARγ)inlungepithelialcellspromotesaPPARγagoNIST-reversIBLeproinflammatoryphenotypeinchronicobstructivepulmonarydisease(COPD)
Authors:SowmyaPLakshmi,AravindTReddy,YingzeZhang,FrankCSciurba,RamaKMallampalli,StevenRDuncan,RajuCReddy
Journal:JournalofBiologicalChemistry(2014):6383--6393
Superoxidedismutaseasatargetofclioquinol-inducedneurotoxicity
Authors:KazuyukiKawamura,YukikoKuroda,MasakoSogo,MikiFujimoto,ToshioInui,TakaoMitsui
Journal:Biochemicalandbiophysicalresearchcommunications(2014):181--185
Xanthineoxidaseinhibitionbyfebuxostatattenuatesexperimentalatherosclerosisinmice
Authors:JohjiNomura,NathalieBusso,AnnetteIves,ChiekoMatsui,SyunsukeTsujimoto,TakashiShirakura,MizuhoTamura,TsunefumiKobayashi,AlexanderSo,YoshihiroYamanaka
Journal:Scientificreports(2014):4554
Highglucose-inducedmitochondrialrespirationandreactiveoxygenspeciesinmousecerebralpericytesisreversedbypharmacologicalinhibitionofmitochondrialcarbonicanhydrases:implicationsforcerebralmicrovasculardiseaseindiabetes
Authors:GulNShah,YoichiMorofuji,WilliamABanks,TulinOPrice
Journal:Biochemicalandbiophysicalresearchcommunications(2013):354--358
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