| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 540/590 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy CellMetabolism |
| Related | RedoxEnzymes |
| Spectrum | AdvancedSpectrumViewer |
- Foreachsample,preparecellsin0.5mLserum-freemediumorbufferofyourchoiceatadensityof1×105to1×106cells/mL.
Note1:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.Foradherentcells,gentlyliftthecellswith0.5mMEDTAtokeepthecellsintact,andwashthecellsoncewithserum-freemedia.
Note2:JZL1707NAD(P)Hsensorisserumsensitive,thereforeit’srecommendedtokeepcellsinserum-freemediumorbufferofyourchoice.Alternatively,cellscanbepreparedandtreatedinregularfullmedium.Changetoserum-freemediumorbufferofyourchoicewhenincubationwithJZL1707NAD(P)Hsensor. - Incubatecellswithtestcompoundsat37ºCforadesiredperiodoftimetostimulateintracellularNADH/NADPH.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandtestcompoundused.Optimizetheincubationtimeforeachexperiment. - Add1μLofJZL1707NAD(P)Hsensor(ComponentA)into0.5mLcellsUSPension.Incubateat37ºCfor30-60minutes.
Note:ForaNADH/NADPHpositivecontroltreatment:Jurkatcellswereincubatedwith100µMNADHorNADPHfor30minutesinserum-freemedium,andco-incubatedwithJZL1707NAD(P)Hsensorworkingsolutionat37ºCforanother30minutes.SeeFigure1fordetails. - Washcellswithyourdesiredbufferonce.KeepcellsinAssayBuffer(ComponentB).
- MonitorthefluorescenceintensityatPEchannelusingaflowcytometer.Gateonthecellsofinterest,excludingdebris.
| References&Citations |  PrinterFriendlyVersion |
1. RubinTan,JianshaLi,XiaochunPeng,LipingZhu,LeiCai,TaoWang,YuanSu,KaikobadIrani,andQinghuaHu.GAPDHiscriticalforsuperiorefficacyoffemalebonemarrow-derivedmesenchymalstemcellsonpulmonaryhypertension.CardiovascRes,Oct2013;100:19-27.
2. StephenY.Xue,ValeriaY.Hebert,DaniciaM.Hayes,CorieN.Robinson,MitziGlover,andTammyR.Dugas.NucleosideReverseTranscriptaseInhibitorsInduceaMitophagy-AssociatedEndothelialCytotoxicityThatIsReversedbyCoenzymeQ10Cotreatment.Toxicol.Sci.,Aug2013;134:323–334.
3. KateJ.Roberts,AndrewCross,OlgaVasieva,RobertJ.Moots,andStevenW.Edwards.Inhibitionofpre-Bcellcolony-enhancingfactor(PBEF/NAMPT/visfatin)decreasestheABIlityofhumanneutrophilstogeneratereactiveoxidantsbutdoesnotimpairbacterialkilling.J.Leukoc.Biol.,Sep2013;94:481-492.
4. WeijingCai,MayaRamdas,LiZhu,XueChen,GaryE.Striker,andHelenVlassara.Oraladvancedglycationendproducts(AGEs)promoteinsulinresistanceanddiabetesbydepletingtheantioxidantdefensesAGEreceptor-1andsirtuin1.PNAS,Sep2012;109:15888-15893.
5. YueQiu,ClausTittiger,ClaudeWicker-Thomas,GaëlleLeGoff,SharonYoung,EricWajnberg,ThierryFricaux,NathalieTaquet,GaryJ.Blomquist,andRenéFeyereisen.Aninsect-specificP450oxidativedecarbonylaseforcuticularhydrocarbonbiosynthesis.PNAS,Sep2012;109:14858-14863.
