Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 447/553 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | CellBIOLOGy LabelingCells |
Related | CellFunctionalAnalysis BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1. Culturecellstoadensityoptimumforautophagyinductionaccordingtoyourspecificinductionprotocol(about1-2×104cells/well/96-wellplate).Atthesametime,cultureanon-inducednegativecontrolcellpopulationatthesamedensityastheinducedpopulationforeverylabelingcondition.
2. PrepareAutophagyGreen™workingsolutionbydiluting20μLofAutophagyGreen™(ComponentA)to10mLofStainBuffer(ComponentB).
Note:20μLof500XAutophagyGreen™(ComponentA)isenoughforone96-wellplate.Aliquotandstoreunused500XAutophagyGreen™at<-20ºC.Protectfromlightandavoidrepeatedfreeze-thawcycles.
3. Removemedium,add100μLofAutophagyGreen™workingsolution(fromStep2)intoeachwell,andincubatethecellsina37ºC,5%CO2incubatorfor15min-1hour.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
4. WashthecellswithWashBuffer(ComponentC)for3-4times,add100μLWashBuffer(ComponentC) toeachwell.
Note:Itisrecommendedtoincreaseeitherthelabelingconcentrationortheincubationtimetoallowthedyetoaccumulateifthecellsdonotappeartobesufficientlystained.
5. MonitorfluorescentintensitywithafluorescencemicroscopeormicroplatereaderatEx/Em=485/530nm.
References&Citations | CitationExplorer |
MethodsforMeasuringAutophagyLevelsinDisease
Authors:KanchanPhadwal,DominicKurian
Journal:(2017):195--211
Highglucoseinducesbonemarrow-derivedmesenchymalstemcellsenescencebyupregulatingautophagy
Authors:Tzu-ChingChang,Min-FenHsu,KennethKWu
Journal:PloSone(2015):e0126537
LicochalconeAinducesautophagythroughPI3K/Akt/mTORinactivationandautophagysuppressionenhancesLicochalconeA-inducedapoptosisofhumancervicalcancercells
Authors:Jen-PiTsai,Chien-HsingLee,Tsung-HoYing,Chu-LiangLin,Chia-LiangLin,Jung-TsungHsueh,Yi-HsienHsieh
Journal:Oncotarget(2015):28851