Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 422/466 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | CellBIOLOGy CellMetabolism |
Related | RedoxEnzymes BiochemicalAssays |
1.PrepareNADstocksolution:
Add500µLofddH2OintothevialofNADstandard(ComponentD)tomake1mMNADstocksolution.
Note:TheunusedNADstocksolutionshouldbedividedintosingleusealiquotsandstoredat-20oC.
2.PrepareserialdilutionsofNADstandard(0to10μM):
2.1 Add10μLofNADstocksolution(fromStep1)into990µLH2OorPBSbuffertogenerate10 µMNADstandardsolution.
Note:DilutedNADstandardsolutionisunstable,andshouldbeusedwithin4hours.
2.2 Take200μLof10μMNADstandardsolutiontoperform1:3serialdilutionsinH2OorPBStoget10,3,1,0.3,0.1,0.03,0.01,and0μMserialdilutionsofNADstandard.
2.3 Add50μLserialdilutionsofNADstandardandNADcontainingtestsamplesintoablack/solidbottom96-wellmicroplateasdescribedinTable1:
Table1LayoutofNADstandardsandtestsamplesinablack/solidbottom96-wellmicroplate
BL | BL | TS | TS | …. | …. |
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NS1 | NS1 | …. | …. | …. | …. |
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NS2 | NS2 |
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NS3 | NS3 |
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NS4 | NS4 |
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NS5 | NS5 |
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NS6 | NS6 |
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NS7 | NS7 |
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Note1:NS=NADStandards,BL=BlankControl,TS=TestSamples.
Note2:AddtheseriallydilutedNADstandardsfrom10μMto0.01μMintowellsfromNS1toNS7induplicate.
3.RunNADassay:
3.1 Add20μLQuestFluor™NADProbe(ComponentA)solutionintoeachwellofNADstandard,blankcontrol,andtestsamples,mixwell.
3.2 Add20μLAssaySolution(ComponentB)intoeachwell,mixwell.
Note:Fora384-wellplate,add25μLofsampleand10μLofQuestFluor™NADProbeand10μLAssaySolutionintoeachwell.
3.3 Incubatethereactionatroomtemperaturefor10-20minutes,protectedfromlight.
3.4 Add15μLEnhancer(ComponentC)toeachwelltomakethetotalNADassayvolumeof105µL/well,andincubateatroomtemperaturefor10-20minutes,protectedfromlight.
Note:Fora384-wellplate,add7.5uLEnhancer.
3.5 Monitorthefluorescenceincreasewithafluorescenceplatereaderat420/480nm.
References&Citations | ![]() CitationExplorer |
CelastrolattenuatesangiotensinIImediatedhumanumbilicalveinendothelialcellsdamagethroughactivationofNrf2/ERK1/2/Nox2signalpathway
Authors:MiaoLi,XinLiu,YongpengHe,QingyinZheng,MinWang,YuWu,YuanpengZhang,ChaoyunWang
Journal:EuropeanJournalofPharmacology(2017):124--133
CytosolicRedoxStatusofWineYeast(SaccharomycesCerevisiae)underHyperosmoticStressduringIcewineFermentation
Authors:FeiYang,CaitlinHeit,DebraLInglis
Journal:Fermentation(2017):61
EpigeneticregulationofRunx2transcriptionandosteoblastdifferentiationbynicotinamidephosphoribosyltransferase
Authors:MinLing,PeixinHuang,ShamimaIslam,DanielPHeruth,XuananLi,LiQinZhang,Ding-YouLi,ZhaohuiHu,ShuiQingYe
Journal:Cell&Bioscience(2017):27
MCU-dependentmitochondrialCa2+inhibitsNAD+/SIRT3/SOD2pathwaytopromoteROSproductionandmetastasisofHCCcells
Authors:TRen,HZhang,JWang,JZhu,MJin,YWu,XGuo,LJi,QHuang,HYang
Journal:Oncogene(2017)
Metabolicandmolecularinsightsintoanessentialroleofnicotinamidephosphoribosyltransferase
Authors:LiQZhang,LeonVanHaandel,MinXiong,PeixinHuang,DanielPHeruth,CharlieBi,RogerGaedigk,XunJiang,Ding-YouLi,GeraldWyckoff
Journal:CellDeath&Disease(2017):e2705
PyrroloquinolineQuinone,aRedox-activeo-Quinone,StimulatesMitochondrialBiogenesisbyActivatingSIRT1/PGC-1αSignalingPathway
Authors:KazuhiroSaihara,RyosukeKamikubo,KazutoIkemoto,KojiUchida,MitsuguAkagawa
Journal:Biochemistry(2017)
ResveratrolattenuatesexcessiveethanolexposureinducedinsulinresistanceinratsviaimprovingNAD+/NADHratio
Authors:GangLuo,BingqingHuang,XiangQiu,LinXiao,NingWang,QinGao,WeiYang,LipingHao
Journal:MolecularNutrition&FoodResearch(2017)
ASnapshotofthePlantGlycatedProteomeSTRUCTURAL,FUNCTIONAL,ANDMECHANISTICASPECTS
Authors:TatianaBilova,ElenaLukasheva,DominicBrauch,UtaGreifenhagen,GaganPaudel,ElenaTarakhovskaya,NadezhdaFrolova,JulianeMittasch,GerdUlrichBalcke,AlainTissier
Journal:JournalofBiologicalChemistry(2016):7621--7636
AMPKactivationprotectscellsfromoxidativestress-inducedsenescenceviaautophagicfluxrestorationandintracellularNAD+elevation
Authors:XiaojuanHan,HaoranTai,XiaoboWang,ZheWang,JiaoZhou,XiaweiWei,YiDing,HuiGong,ChunfenMo,JieZhang
Journal:Agingcell(2016):416--427
Cell-LineSelectivityImprovesthePredictivePowerofPharmacogenomicAnalysesandHelpsIdentifyNADPHasBioMarkerforFerroptosisSensitivity
Authors:KenichiShimada,MikiHayano,NenCPagano,BrentRStockwell
Journal:Cellchemicalbiology(2016):225--235
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