>
产品中心 >
Other_kits >
AAT Bioquest/Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Green Fluorescence*/22900/200 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 492/520 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | NeuroBIOLOGy ReactiveOxygenSpecies |
| Related | CellSignaling CellFunctionalAnalysis BiochemicalAssays |
1.Preparecells:
1.1 Foradherentcells:Platecellsovernightingrowthmediumat10,000to40,000cells/well/100µLfora96-wellplateor2,500to10,000cells/well/25µLfora384-wellplate.
1.2 Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletsinculturemediumat50,000-100,000cells/well/100µLfora96-wellpoly-Dlysineplateor10,000-25,000cells/well/25µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiment.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.
2.PrepareAmplite™ROSGreenstainsolution:
2.1 PrepareAmplite™ROSGreenstocksolution(500X):Add40µLofDMSO(ComponentC)intothevialofAmplite™ROSGreen(ComponentA),andmixthemwell.
Note:20µLofAmplite™ROSGreenstocksolutionisenoughfor1plate.Un-usedreconstitutedAmplite™ROSGreenstocksolutioncanbealiquotedandstoredat<-20°Cformorethanonemonthifthetubesaresealedtightlyandprotectedfromlight.Avoidrepeatedfreeze-thawcycles.
2.2 PrepareAmplite™ROSGreenworkingsolution:Add20µLof500XDMSOreconstitutedAmplite™ROSGreenstocksolution(fromStep2.1)into10mLofAssayBuffer(ComponentB),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.
3.Stainthecells:
3.1 Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofAmplite™ROSGreenworkingsolution(fromStep2.2)intothecellplate.
3.2 Incubatethecellsina5%CO2,37°Cincubatorforonehour.
4.RunROSAssay:
4.1 Treatcellsbyadding20μL/wellof11Xtestcompounds(96-wellplate)or10μL/wellof6Xtestcompounds(384-wellplate)inthedesiredbuffer(suchasPBSorHHBS).Forcontrolwells(untreatedcells),addthecorrespondingamountofcompoundbuffer.
4.2 Incubatethecellplateatroomtemperatureorina5%CO2,37°C,incubatorforatleast15minutesoradesiredperiodoftime(30minutesforHelacellstreatedwith1mMH2O2)toinduceROS.
4.3 MonitorthefluorescenceincreaseatEx/Em=490/525nmwithbottomreadmode.
| 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
