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AAT Bioquest/Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Blue Fluorescence Excited at 405 nm*/22828/100 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 405/450 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.PrepareandincubatecellswithAnnexinV-mFluorViolet™450:
1.1 Treatcellswithtestcompoundsforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithstaurosporine)toinduceapoptosis.
1.2 Centrifugethecellstoget1-5×105cells/tube.
1.3 ResUSPendcellsin200μLofAssayBuffer(ComponentB).
1.4 Add2μLofAnnexinV-mFluorViolet™450(ComponentA)intothecells.
Optional:Add2µLof100XPropidiumIodide(ComponentC)intothecellsfornecrosiscells.
1.5 Incubateatroomtemperaturefor30to60minutes,protectedfromlight.
1.6 Add300μLofAssayBuffer(ComponentB)toincreasevolumebeforeanalyzingthecellswithaflowcytometerorfluorescencemicroscope(seeStep1.7below).
1.7 MonitorthefluorescenceintensityatEx/Em=405/450nmbyusingaflowcytometerorafluorescencemicroscope(SeeStep2or3below).
2.Analyzebyusingaflowcytometer:
QuantifyAnnexinV-mFluorViolet™450bindingbyusingaflowcytometeratEx/Em=405/450nm.MeasurethecellviabilitybyusingtheFL2channelwhenpropidiumiodideisaddedintothecells.
Note:AnnexinVbindingflowcytometricanalysisonadherentcellsisnotroutinelytestedsincespecificmembranedamagemayoccurduringcelldetachmentorharvesting.However,methodsforutilizingAnnexinVforflowcytometryonadherentcelltypeshavebeenpreviouslyreportedbyCasiola-Rosenetal.andvanEngelendetal(seeRefs1and2).
3.Analyzebyusingafluorescencemicroscope:
3.1 PipettethecellsuspensionfromStep1.5,rinse1-2timeswithassaybuffer,andthenresuspendthecellswithassaybuffer.Addthecellsonaglassslidethatiscoveredwithaglasscoverslip.
Note:Foradherentcells,itisrecommendedtogrowthecellsdirectlyonacoverslip.AfterincubationwithAnnexinV-mFluorViolet™450(Step1.5),rinse1-2timeswithassaybuffer,andaddassaybufferbacktothecoverslip.Invertcoversliponaglassslideandvisualizethecells.Thecellscanalsobefixedin2%formaldehydeaftertheincubationwithAnnexinV-mFluorViolet™450andvisualizedunderamicroscope.
3.2 AnalyzetheapoptoticcellswithAnnexinV-mFluorViolet™450underafluorescencemicroscopeusingtheVioletchannel.MeasurethecellviabilitybyusingtheTRITCchannelwhenpropidiumiodideisaddedintothecells.ThebluestainingontheplasmamembraneindicatestheAnnexinV-mFluorViolet™450bindingtoPSoncellsurface.
| References&Citations |  CitationExplorer |
Insitupolymerizablehydrogelincorporatedwithspecificpathogen-freeporcineplatelet-richplasmaforthereconstructionofthecornealendothelium
Authors:Yung-KaiLin,RuchiSharma,HsuMa,Wen-ShyanChen,Chao-LingYao
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NovelregulationsofMEF2-A,MEF2-D,andCACNA1SinthefunctionalincompetenceofADIpose-derivedmesenchymalstemcellsbyinducedindoxylsulfateinchronickidneydisease
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Journal:Cytotechnology(2016):2589--2604
Shearstress-inducedalterationofepithelialorganizationinhumanrenaltubularcells
Authors:DamienMaggiorani,RomainDissard,MarcyBelloy,Jean-SébastienSaulnier-Blache,AudreyCasemayou,LaureDucasse,SandraGrès,JulieBellière,CécileCaubet,Jean-LoupBascands
Journal:PloSone(2015):e0131416
TargetingaG-protein-coupledreceptoroverexpressedinendocrinetumorsbymagneticnanoparticlestoinducecelldeath
Authors:ClaireSanchez,DarineElHajjDiab,VincentConnord,PascalClerc,EtienneMeunier,BernardPipy,BrunoPayré,ReasmeyPTan,MichelGougeon,JulianCarrey
Journal:AcsNano(2014):1350--1363
