| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 445/503 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy LabelingCells |
| Related | FluorescenceImaging BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.Preparelysosomal-stainingsolution:
1.1 WarmLysolite™Green(ComponentA)toroomtemperature.
1.2 Preparedyeworkingsolutionbydiluting20µLofLysolite™Green(ComponentA)to10mLofLiveCellStainingBuffer(ComponentB).
Note1:20µLofLysolite™Green(ComponentA)isenoughforone96-wellplate.AliquotandstoreunusedLysolite™Green(ComponentA)at<‑20oC.Protectfromlightandavoidrepeatedfreeze-thawcycles.
Note2:Theoptimalconcentrationofthefluorescentlysosomeindicatorvariesdependingonthespecificapplication.Thestainingconditionsmaybemodifiedaccordingtotheparticularcelltypeandthepermeabilityofthecellsortissuestotheprobe.
2.Prepareandstaincells:
2.1 Foradherentcells:Growcellseitherina96-wellblackwall/clearbottomplate(100µL/well/96-wellplate)oroncover-slipsinsideapetridishfilledwiththeappropriateculturemedium.Whencellsreachthedesiredconfluence,addequalvolume(suchas100µL/well/96-wellplate)ofthedye-workingsolution(fromStep1.2).Incubatethecellsina37°C,5%CO2incubatorfor30minutesto2hours.ObservethecellsusingafluorescencemicroscopefittedwithaFITCfilterset.
Note:Itisrecommendedtoincreaseeitherthelabelingconcentrationortheincubationtimetoallowthedyetoaccumulateifthecellsdonotappeartobesufficientlystained.
2.2 Forsuspensioncells:Centrifugethecellsat1000rpmfor5minutestoobtainacellpelletandaspiratethesupernatant.Resuspendthecellpelletgentlyinpre-warmedgrowthmedium,andthenaddequalvolumeofthedye-workingsolution(fromStep1.2).Incubatethecellsina37°C,5%CO2incubatorfor30minutesto2hours.ObservethecellsusingafluorescencemicroscopefittedwithaFITCfilterset.
Note1:Itisrecommendedtoincreaseeitherthelabelingconcentrationortheincubationtimetoallowthedyetoaccumulateifthecellsdonotappeartobesufficientlystained.
Note2:Suspensioncellsmaybeattachedtocover-slipsthathavebeentreatedwithBDCell-Tak®(BDBiosciences)andstainedasadherentcells(seeStep2.1).
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