| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 646/659 |
| MW | ~350 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | CellBIOLOGy CellFunctionalAnalysis |
| Related | HydrolyticEnzymes BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.Preparecells:
Plate100to100,000x10cellsperwellinatissueculturemicroplatewithblackwallandclearbottom.Addtestcompoundsintothecellsandincubateforadesiredperiodoftime(suchas24,48or96hours)ina37°C,5%CO2incubator.Forblankwells(mediumwithoutthecells),addthesameamountofcompoundbuffer.Thesuggestedtotalvolumeis100µL/well/96-wellplate,and25µL/well/384-wellplate.
Note:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensityforproliferationorcytotoxicityinduction.Forproliferationassays,usefewercells;forcytotoxicityassays,usemorecellstostartwith.
2.PrepareSunRed™acetateworkingsolution:
2.1 Preparea2to10mMstocksolutionofSunRed™acetateinDMSO.Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandfrozenat<-20oC.Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
2.2 PrepareSunRed™acetateworkingsolutionat5to10µMin1XHank"ssaltsolutionwith20mMHepesbuffer(HHBS)orbufferofyourchoicebeforetheexperiment.
3.Runthecellviabilityassay:
3.1 Treatcellswithtestcompoundsasdesired(fromStep1).
Note:Itisnotnecessarytowashcellsbeforeaddingcompound.However,iftestedcompoundsareserumsensitive,growthmediumandserumfactorscanbeaspiratedawaybeforeaddingcompounds.Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofHHBSorthebufferofyourchoiceafteraspiration.Alternatively,cellscanbegrowninserum-freemedia.
3.2 Removethemediumfromthecells.
3.3 Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofSunRed™acetateworkingsolution(fromStep2.2).
3.4 IncubatetheSunRed™acetateworkingsolutionplateatroomtemperatureor37oCfor1hour,protectedfromlight.
Note1:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
Note2:Fornon-adherentcells,itisrecommendedtocentrifugecellplatesat800rpmfor2minuteswithbrakeoffafterincubation.
3.5 MonitorthefluorescenceintensityatEx/Em=620/660nm(cutoff640nm)
| References&Citations |  PrinterFriendlyVersion |
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