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AAT Bioquest/Cell Meter™ Fluorimetric Cell Cycle Assay Kit *Optimized for 405 nm Violet Laser Excitation*/22845/100 Te
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 401/459 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1. Foreachsample,preparecellsin0.5mLofwarmmediumorbufferofyourchoiceatadensityof5×105to1×106cells/mL.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforapoptosisinduction.
2. Treatcellswithtestcompoundsforadesiredperiodoftimetoinduceapoptosisorothercellcyclefunctions.
3. Add1μLof500XNuclearViolet™(ComponentA)intothecellscontainingthegrowthmedium,andincubatethecellsina37°C,5%CO2incubatorfor30to60minutes.
Note1:Foradherentcells,gentlyliftthecellswith0.5mMEDTAtokeepthecellsintact,andwashthecellsoncewithserum-containingmediapriortoincubationwithNuclearViolet™.
Note2:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
Note3:ItisnotnecessarytofixthecellsbeforeDNAstainingsincetheNuclearViolet™iscell-permeable.
Note4:HumanandrodentstemcellseffluxNuclearViolet™stain,andthisisthebasisfortheSidePopulation(SP)technique;theeffluxcanbeblockedwithblockingagentslikeverapamil,fumitermorginCtopreventdyeeffluxforaccurateDNAcontentanalysis.
4. Optional:Centrifugethecellsat1000rpmfor4minutes,andthenre-sUSPendcellsin0.5mLofassaybuffer(ComponentB)orthebufferofyourchoice.
5. Monitorthefluorescenceintensitybyflowcytometryusingthe405nmvioletlaser(Ex/Em=405/445nm).Gateonthecellsofinterest,excludingdebris.
| References&Citations |  PrinterFriendlyVersion |
2. EngstromJU,KmiecEB.(2008)DNAreplication,cellcycleprogressionandthetargetedgenerepairreaction.CellCycle,7,1402.
3. KuoHM,ChangLS,LinYL,LuHF,YangJS,LeeJH,ChungJG.(2007)MorininhibitsthegrowthofhumanleukemiaHL-60cellsviacellcyclearrestandinductionofapoptosisthroughmitochondriadependentpathway.AnticancerRes,27,395.
4. LouieMC,RevenkoAS,ZouJX,YaoJ,ChenHW.(2006)Directcontrolofcellcyclegeneexpressionbyproto-oncogeneproductACTR,anditsautoregulationunderliesitstransformingactivity.MolCellBiol,26,3810.
5. EaswaranHP,LeonhardtH,CardosoMC.(2005)CellcycleMarkersforlivecellanalyses.CellCycle,4,453.
6. EssersJ,vanCappellenWA,TheilAF,vanDrunenE,JaspersNG,HoeijmakersJH,WymanC,VermeulenW,KanaarR.(2005)Dynamicsofrelativechromosomepositionduringthecellcycle.MolBiolCell,16,769.
7. LunaL,RolsethV,HildrestrandGA,OtterleiM,DantzerF,BjorasM,SeebergE.(2005)DynamicrelocalizationofhOGG1duringthecellcycleisdisruptedincellsharbouringthehOGG1-Cys326polymorphicvariant.NucleicAcidsRes,33,1813.
8. Baran-MarszakF,FeuillardJ,NajjarI,LeClorennecC,BechetJM,Dusanter-FourtI,BornkammGW,RaphaelM,FagardR.(2004)DifferentialrolesofSTAT1alphaandSTAT1betainfludarABIne-inducedcellcyclearrestandapoptosisinhumanBcells.Blood,104,2475.
9. ConlonKA,ZharkovDO,BerriosM.(2004)Cellcycleregulationofthemurine8oxoguanineDNAglycosylase(mOGG1):mOGG1associateswithmicrotubulesduringinterphaseandmitosis.DNARepair(Amst),3,1601.
10. NunezR,GarayN,VillafaneC,BrunoA,LindgrenV.(2004)DescriptionofaflowcytometryapproachbasedonSYBR-14stainingfortheassessmentofDNAcontent,cellcycleanalysis,andsortingoflivingnormalandneoplasticcells.ExpMolPathol,76,29.
