Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 492/519 |
MW | ~500 |
CAS# | N/A |
Solvent | DMSO |
Storage | F/D/L |
Category | CellBIOLOGy LabelingCells |
Related | FluorescenceImaging ViABIlityandProliferation BiochemicalAssays |
1.Prepare500XDMSOstocksolution
Add500µLDMSOintothedyepowdervial,mixitwellbyvortexingtohavea500XDMSOstocksolution
Note:Thestocksolutionshouldbeusedpromptly;anyremainingsolutionshouldbealiquotedandfrozenat<-20oC.Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
2.Prepare1Xdyeworkingsolution
Preparea1Xdyeworkingsolutionbydilutingthe500XDMSOstocksolutionat1to500 inHanksand20mMHepesbuffer(HHBS)orthebufferofyourchoice,pH7(suchas1µLof500XDMSOstocksolutionto500µLbuffer) rightbeforeuse.Mixthemwellbyvortexing.
Note:Thefinalconcentrationofthedyeworkingsolutionshouldbeempiricallydeterminedfordifferentcelltypesand/orexperimentalconditions.Itisrecommendedtotestattheconcentrationsthatareatleastoveratenfoldrange.SuchasCytoTell™Redmightusemuchlessamountinsomecelltypesthantherecommendconcentrations.
3.Analyzecellswithaflowcytometerorafluorescencemicroscope:
3.1 Treatcellswithtestcompoundsforadesiredperiodoftime.
3.2 Centrifugethecellstoget1-5×105cellspertube.
3.3 ResUSPendcellsin500μLofthedyeworkingsolution(fromStep2).
Optional:Onecanaddthe500XDMSOstocksolutionintothecellsdirectlywithoutmediumremoving(suchas,add1µL500XDMSOstocksolutioninto500µLcells)
3.4 Incubatecellswithadyesolutionatroomtemperatureor37°Cfor10to30min,protectedfromlight.
3.5 Removethedyeworkingsolutionfromthecells,washthecellswithHHBSorbufferofyourchoice.Resuspendcellsin500μLofpre-warmedHHBSormediumtoget1-5×105cellspertube.
3.6 MonitorthefluorescencechangeatrespectedEx/Em(seeTable1)withaflowcytometerorafluorescencemicroscope.
References&Citations | CitationExplorer |
CooperationofinnateimmunecellsduringHepatitisCvirusinfection
Authors:VolkerKlöss
Journal:(2017)
CXCL12--CXCR4AxisIsRequiredforContact-MediatedHumanBLymphoidandPlasmacytoidDendriticCellDifferentiationbutNotTLymphoidGeneration
Authors:HirohitoMinami,KeikiNagaharu,YoshikiNakamori,KohshiOhishi,NaoshiShimojo,YukiKageyama,TakeshiMatsumoto,YukaSugimoto,IsaoTawara,MasahiroMasuya
Journal:TheJournalofImmunology(2017):ji1700054
InteractionandMutualActivationofDifferentInnateImmuneCellsIsNecessarytoKillandClearHepatitisCVirus-InfectedCells
Authors:VolkerKlöss,OliverGrünvogel,GuidoWabnitz,TatjanaEigenbrod,StefanieEhrhardt,FelixLasitschka,VolkerLohmann,AlexanderHDalpke
Journal:FrontiersinImmunology(2017):1238
interactionandMutualactivationofDifferentinnateimmunecellsisnecessarytoKillandclearhepatitiscVirus-infectedcells
Authors:VolkerKlöss,OliverGrünvogel,GuidoWabnitz,TatjanaEigenbrod,StefanieEhrhardt,FelixLasitschka,VolkerLohmann,AlexanderHDalpke
Journal:FrontiersinImmunology(2017):1238
OnioninAinhibitsovariancancerprogressionbysuppressingcancercellproliferationandtheprotumourfunctionofmacrophages
Authors:JunkoTsuboki,YukioFujiwara,HasitaHorlad,DaisukeShiraishi,ToshihiroNohara,ShingoTayama,TakeshiMotohara,YoichiSaito,TsuyoshiIkeda,KiyomiTakaishi
Journal:ScientificReports(2016)
Multiplexinganalysisofcellproliferationandcellularfunctionsusinganewmulticolorpaneloffluorescentcellproliferationdyes(P1290)
Authors:JinfangLiao,QinZhao,YiboWu,ZhenjunDiwu
Journal:TheJournalofImmunology(2013):119--4