Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 333/518 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | CellBIOLOGy LabelingCells |
Related | CellFunctionalAnalysis BiochemicalAssays |
1. Culturecellstoadensityoptimumforautophagyinductionaccordingtoyourspecificinductionprotocol(about1-2x104cells/well/96-wellplate).Atthesametime,cultureanon-inducednegativecontrolcellpopulationatthesamedensityastheinducedpopulationforeverylabelingcondition.
2. PrepareAutophagyBlue™workingsolutionbydiluting20μLofAutophagyBlue™(ComponentA)to10mLofStainBuffer(ComponentB).
Note:20μLof500XAutophagyBlue™(ComponentA)isenoughforone96-wellplate.Aliquotandstoreunused500XAutophagyBlue™at<-20°C.Protectfromlightandavoidrepeatedfreeze-thawcycles.
3. Removemedium,add100μLofAutophagyBlue™workingsolution(fromStep2)intoeachwell,andincubatethecellsina37°C,5%CO2incubatorfor15min-1hour.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
4. WashthecellswithWashBuffer(ComponentC)for3-4times,add100μLWashBuffer(ComponentC) toeachwell.
Note:Itisrecommendedtoincreaseeitherthelabelingconcentrationortheincubationtimetoallowthedyetoaccumulateifthecellsdonotappeartobesufficientlystained.
5. Monitorthefluorescentintensitywithafluorescencemicroscope,flowcytometer,orfluorescencemicroplatereaderatEx/Em=330/520nm.
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