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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Red Fluorescence*/22901/200 Tests
产品编号:22901
市  场 价:¥56560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$2828.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Red Fluorescence*/22901/200 Tests
商品介绍
Overview
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Ex/Em(nm)520/605
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryNeuroBIOLOGy
ReactiveOxygenSpecies
RelatedCellSignaling
CellFunctionalAnalysis
BiochemicalAssays
Reactiveoxygenspecies(ROS)arenaturalbyproductsofthenormalmetabolismofoxygenandplayimportantrolesincellsignaling.However,duringoxidativestress-relatedstates,ROSlevelscanincreasedramatically.TheaccumulationofROSresultsinsignificantdamagetocellstructures.Theroleofoxidativestressincardiovasculardisease,diabetes,osteoporosis,stroke,inflammatorydiseases,anumberofneurodegenerativediseasesandcancerhasbeenwellestablished.TheROSmeasurementwillhelptodeterminehowoxidativestressmodulatesvariedintracellularpathways.Amplite™FluorimetricROSAssayKitusesouruniqueROSsensortoquantifyROSinlivecells.Amplite™ROSRediscell-permeable.ItgeneratestheredfluorescencewhenitreactswithROS.Thekitisanoptimized"mixandread"assayformat.TheAmplite™FluorimetricROSAssayKitprovidesasensitive,one-stepfluorimetricassaytodetectintracellularROSinlivecellswith1-2hoursincubation.Theassaycanbeperformedinaconvenient96-wellor384-wellmicrotiter-plateformatandeasilyadaptedtoautomationwithoutaseparationstep.ItssignalcanbeeasilyreadusingeitherafluorescencemicroplatereaderorafluorescentmicroscopeatEx/Em=520/605nm.ItcanbeusedtoeitherquantifytheROSactivitiesorscreentheROSinhibitors.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecells:

1.1   Foradherentcells:Platecellsovernightingrowthmediumat10,000to40,000cells/well/100µLfora96-wellplateor2,500to10,000cells/well/25µLfora384-wellplate.

 

1.2   Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandsUSPendthecellpelletsinculturemediumat50,000-100,000cells/well/100µLfora96-wellpoly-Dlysineplateor10,000-25,000cells/well/25µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortoyourexperiment.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.

2.PrepareAmplite™ROSRedstainsolution:

2.1   PrepareAmplite™ROSRedstocksolution(500X):Add40µLofDMSO(ComponentC)intothevialofAmplite™ROSRed(ComponentA),andmixthemwell.

Note:20µLofreconstitutedAmplite™ROSRedstocksolutionisenoughfor1plate.Unusedportioncanbealiquotedandstoredat<-20°Cformorethanonemonthifthetubesaresealedtightlyandkeptfromlight.Avoidrepeatedfreeze-thawcycles.

 

2.2   PrepareAmplite™ROSRedworkingsolution:Add20µLof500XDMSOreconstitutedAmplite™ROSRedstocksolution(fromStep2.1)into10mLofAssayBuffer(ComponentB),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.

  

3.Stainthecells:

3.1   Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofAmplite™ROSRedworkingsolution(fromStep2.2)intothecellplate.

3.2   Incubatethecellsina5%CO2,37°Cincubatorforonehour.

 

4.RunROSAssay:

4.1   Treatcellswith20μLof11Xtestcompounds(96-wellplate)or10μLof6Xtestcompounds(384-wellplate)inyourdesiredbuffer(suchasPBSorHHBS).Forcontrolwells(untreatedcells),addthecorrespondingamountofcompoundbuffer.

4.2   ToinduceROS,incubatethecellplateatroomtemperatureorina5%CO2,37°Cincubatorforatleast15minutesoradesiredperiodoftime(30minutesforHelacellstreatedwith1mMH2O2).

4.3   MonitorthefluorescenceincreaseatEx/Em=520/605nmwithbottomreadmode.

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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