| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 554/578 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.PrepareandincubatecellswithAnnexinV-iFluor™555:
1.1 Treatcellswithtestcompoundsforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithstaurosporine)toinduceapoptosis.
1.2 Centrifugethecellstoget1-5×105cells/tube.
1.3 ResUSPendcellsin200μLofAssayBuffer(ComponentB).
1.4 Add2μLofAnnexinV-iFluor™555(ComponentA)intothecells.
1.5 Incubateatroomtemperaturefor30to60minutes,protectedfromlight.
1.6 Add300μLofAssayBuffer(ComponentB)toincreasevolumebeforeanalyzingthecellswithaflowcytometerorfluorescencemicroscope(seeStep1.7below).
1.7 MonitorthefluorescenceintensityatEx/Em=555/590nmbyusingaflowcytometerorafluorescencemicroscope(SeeStep2or3below).
2.Analyzebyusingaflowcytometer:
QuantifyAnnexinV-iFluor™555bindingbyusingaflowcytometeratEx/Em=555/590nm.
Note:AnnexinVbindingflowcytometricanalysisonadherentcellsisnotroutinelytestedsincespecificmembranedamagemayoccurduringcelldetachmentorharvesting.However,methodsforutilizingAnnexinVforflowcytometryonadherentcelltypeshavebeenpreviouslyreportedbyCasiola-Rosenetal.andvanEngelendetal(seeRefs1and2).
3.Analyzebyusingafluorescencemicroscope:
3.1 PipettethecellsuspensionfromStep1.5,rinse1-2timeswithassaybuffer,andthenresuspendthecellswithassaybuffer.Addthecellsonaglassslidethatiscoveredwithaglasscoverslip.
Note:Foradherentcells,itisrecommendedtogrowthecellsdirectlyonacoverslip.AfterincubationwithAnnexinV-iFluor™555(Step1.5),rinse1-2timeswithassaybuffer,andaddassaybufferbacktothecoverslip.Invertcoversliponaglassslideandvisualizethecells.Thecellscanalsobefixedin2%formaldehydeaftertheincubationwithAnnexinV-iFluor™555andvisualizedunderamicroscope.
3.2 AnalyzetheapoptoticcellswithAnnexinV-iFluor™555underafluorescencemicroscopeusingtheVioletchannel.MeasurethecellviabilitybyusingtheCy5channelwhenNuclearRed™DCS1(AATcatalog#17552)isaddedintothecells.TheorangestainingontheplasmamembraneindicatestheAnnexinV-iFluor™555bindingtoPSoncellsurface.
| References&Citations |  CitationExplorer |
Insitupolymerizablehydrogelincorporatedwithspecificpathogen-freeporcineplatelet-richplasmaforthereconstructionofthecornealendothelium
Authors:Yung-KaiLin,RuchiSharma,HsuMa,Wen-ShyanChen,Chao-LingYao
Journal:JournaloftheTaiwanInstituteofChemicalEngineers(2017)
NovelregulationsofMEF2-A,MEF2-D,andCACNA1SinthefunctionalincompetenceofADIpose-derivedmesenchymalstemcellsbyinducedindoxylsulfateinchronickidneydisease
Authors:DuyenThiDo,NamNhutPhan,Chih-YangWang,ZhengdaSun,Yen-ChangLin
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
