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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*Optimized for Flow Cytometry*/22904/1
产品编号:22904
市  场 价:¥85560.00
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:$4278.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Optimized for Flow Cytometry*/22904/1
商品介绍
Overview
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Ex/Em(nm)490/520
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.CellMeter™FluorimetricROSAssayKitusesouruniqueAmplite™ROSGreensensortoquantifyROSinlivecells.Amplite™ROSGreeniscell-permeable.ItgeneratesthegreenfluorescencewhenitreactswithROS,andcanbeeasilyreadatEx/Em=490/520nm.TheCellMeter™FluorimetricROSAssayKitprovidesasensitive,one-stepfluorimetricassaytodetectintracellularROSinlivecellswithonehourincubation.Thiskitisoptimizedforflowcytometryapplications,itssignalcanbedetectedwithEx/Em=490/520nm(FL1channel).

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.      Foreachsample,preparecellsin0.5mLAssayBuffer(ComponentB)orbufferofyourchoiceatadensityof5×105to1×106cells/mL.

Note:EachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforROSinduction.

 

2.      PrepareAmplite™ROSGreenstocksolution(500X):Add100μLofDMSO(ComponentC)intothevialofAmplite™ROSGreen(ComponentA),andmixwell.

Note:UnusedreconstitutedAmplite™ROSGreenstocksolutioncanbealiquotedandstoredat<-20°Cformorethanonemonthifthetubesaresealedtightlyandkeptfromlight.Avoidrepeatedfreeze-thawcycles.

 

3.      Add1μLof500XAmplite™ROSGreen(ComponentA)into0.5mLcellsUSPension,andincubateat37ºCfor1hour.

Note1:Foradherentcells,gentlyliftthecellswith0.5mMEDTAtokeepthecellsintact,andwashthecellsoncewithserum-containingmediapriortoincubationwithAmplite™ROSGreen.

Note2:Theappropriateincubationtimedependsontheindividualcelltypeandtestcompoundused.Optimizetheincubationtimeforeachexperiment.

 

4.      Treatcellsbyadding50μLof11Xtestcompoundsinthedesiredbuffer(suchasPBSorHBSS).Forcontrolwells(untreatedcells),addthecorrespondingamountofbuffer.

 

5.      Incubatethecellsat37ºCforadesiredperiodoftimetoinduceROS,protectedfromlight.

Note:WetreatedJurkatcellswith100µMTBHP(tert-Butylhydroperoxide)at37ºCfor30minutestoinduceROS.SeeFigure1fordetails.

 

6.      MonitorthefluorescenceintensityattheFL1channel(Ex/Em=490/520nm)usingaflowcytometer.Gateonthecellsofinterest,excludingdebris.

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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