Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 560/574 |
MW | 1015.79 |
CAS# | N/A |
Solvent | DMSO |
Storage | F/D/L |
Category | CellBIOLOGy LabelingCells |
Related | ViABIlityandProliferation CellViability MDRResearch BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
AMestersarethenon-polarestersthatreADIlycrosslivecellmembranes,andrapidlyhydrolyzedbycellularesterasesinsidelivecells.AMestersarewidelyusedforloadingavarietyofpolarfluorescentprobesintolivecellsnon-invasively.However,cautionsmustbeexcisedwhenAMestersareusedsincetheyaresusceptibletohydrolysis,particularlyinsolution.Theyshouldbereconstitutedjustbeforeuseinhigh-quality,anhydrousdimethylsulfoxide(DMSO).DMSOstocksolutionsmaybestoreddesiccatedat‑20°Candprotectedfromlight.Undertheseconditions,AMestersshouldbestableforseveralmonths.
FollowingisourrecommendedprotocolforloadingAMestersintolivecells.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
a) Preparea2to5mMstocksolutionofAMestersinhigh-quality,anhydrousDMSO.ThenonionicdetergentPluronic®F-127issometimesusedtoincreasetheaqueoussolubilityofAMesters.
Note:Anequalvolumeof20%Pluronic®F-127solutioncanbeaddedtoDMSOstocksolutionsofcellviabilityindicatorbeforedilutingintotheloadingbuffer.ThefinalPluronic®F-127concentrationisabout0.02%.AvarietyofPluronic®F-127solutionscanbepurchasedfromAATBioquest.
Caution:Thelong-termstorageofAMestersinthepresenceofPluronic®F-127isnotrecommended.
b) Onthedayoftheexperiment,eitherdissolvethesolidindicatorsofcellviabilityinDMSOorthawanaliquotoftheindicatorstocksolutionstoroomtemperature.Prepareaworkingsolutionof1to10µMinthebufferofyourchoice(suchasHanksandHepesbuffer).Formostofthecelllines,cellviabilityindicatorsatthefinalconcentrationrangingfrom4to5µMarerecommended.Theexactconcentrationofindicatorsrequiredforcellloadingmustbedeterminedempirically.
c) Ifyourcellscontainingtheorganicanion-transports,probenecid(1–2.5mM)orsulfinpyrazone(0.1–0.25mM)maybeaddedtothecellmediumtoreduceleakageofthede-esterifiedindicators.IncubatecellswithAMestersatroomtemperatureor37°Cfor20minutestoonehour.
Note:Decreasingtheloadingtemperaturemightreducetheindicatorcompartmentalization.
d) Washcellsincellviabilityindicator-freebuffer(containingananiontransporterinhibitor,ifapplicable)toremoveexcessprobes.
Note:Cellsampleswereimmediatelyanalyzedbyflowcytometrytodeterminetheaveragefluorescencepercellattimezerowithoutwashing.
e) RuntheexperimentsatthedesiredEx/Emwavelengths(seeTable1).
References&Citations | ![]() CitationExplorer |
Functionalevidencethattheself-renewalgeneNANOGregulatesesophagealsquamouscancerdevelopment
Authors:DengLi,XiaocongXiang,FeiYang,DongqinXiao,KangLiu,ZhuChen,RuolanZhang,GangFeng
Journal:BiochemicalandBiophysicalResearchCommunications(2017)
NINJ2--Anovelregulatorofendothelialinflammationandactivation
Authors:JingjingWang,JingjingFa,PengyunWang,XinzhenJia,HuixinPeng,JingChen,YifanWang,ChenhuiWang,QiuyunChen,XinTu
Journal:CellularSignalling(2017)
ErythropoietinStimulatesEndothelialProgenitorCellstoInduceEndothelializationinanAneurysmNeckAfterCoilEmbolizationbyModulatingVascularEndothelialGrowthFactor
Authors:PeixiLiu,YingjieZhou,QingzhuAn,YayingSong,XiChen,Guo-YuanYang,WeiZhu
Journal:MEDICINE(2016):1--8
FlexibleEndoscopicSprayApplicationofRespiratoryEpithelialCellsasPlatformTechnologytoApplyCellsinTubularOrgans
Authors:AnjaLenaThiebes,ManuelArminReddemann,JohannesPalmer,ReinholdKneer,StefanJockenhoevel,ChristianGabrielCornelissen
Journal:TissueEngineeringPartC:Methods(2016):322--331
Influenceofhypothermiaandsubsequentrewarminguponleukocyte-endothelialinteractionsandexpressionofJunctional-Adhesion-MoleculesAandB
Authors:NicolaiVBogert,IsabellaWerner,AngelaKornberger,PatrickMeybohm,AntonMoritz,TillKeller,UlrichAStock,AndresBeiras-Fernandez
Journal:Scientificreports(2016)
Influenceofhypothermiaandsubsequentrewarminguponleukocyte-endothelialinteractionsandexpressionofJunctional-Adhesion-MoleculesAandB
Authors:NicolaiVBogert,IsabellaWerner,AngelaKornberger,PatrickMeybohm,AntonMoritz,TillKeller,UlrichAStock,AndresBeiras-Fernandez
Journal:Scientificreports(2016)
InhibitionofABCtransportproteinsbyoilsandsprocessaffectedwater
Authors:HattanAAlharbi,DavidMVSaunders,AhmedAl-Mousa,JaneAlcorn,AlbertoSPereira,JonathanWMartin,JohnPGiesy,SteveBWiseman
Journal:AquaticToxicology(2016):81--88
Rapidgenerationofcollagen-basedmicrotissuestostudycell--matrixinteractions
Authors:Marie-ElenaBrett,AlexandraLCrampton,DavidKWood
Journal:Technology(2016):1--8
ToxicokineticsandtoxicodynamicsofchlorpyrifosisalteredinembryosofJapanesemedakaexposedtooilsandsprocess-affectedwater:evidenceforinhibitionofP-glycoprotein
Authors:HattanAAlharbi,JaneAlcorn,AhmedAl-Mousa,JohnPGiesy,SteveBWiseman
Journal:JournalofAppliedToxicology(2016)
Sprayingrespiratoryepithelialcellstocoattissue-engineeredconstructs
Authors:AnjaLenaThiebes,StefanieAlbers,ChristianKlopsch,StefanJockenhoevel,ChristianGCornelissen
Journal:BioResearchopenaccess(2015):278--287
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