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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Calcein Deep Red™ acetate/22010/1 mg
产品编号:22010
市  场 价:¥3900.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$195.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Calcein Deep Red™ acetate/22010/1 mg
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)646/659
MW~500
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryCellBIOLOGy
LabelingCells
RelatedViABIlityandProliferation
CellViability
MDRResearch
BiochemicalAssays
CalceinAMisonethemostpopularfluorescentprobesusedforlabelingandmonitoringcellularfunctionsoflivecells.However,thesinglecolorofCalceinAMmakesitimpossIBLetousethisvaluablereagentinthemulticolorapplications.Forexample,itisimpossibletouseCalceinAMincombinationofGFP-tranfactedcellsduetothesamecolortoGFP.ToaddressthiscolorlimitationofCalceinAM,wehavedevelopedCalceinOrange™,CalceinRed™andCalceinDeepRed™.ThesenewCalceinAManalogsenablethemulticolorlabelingandfunctionalanalysisoflivecellsincombinationwithCalceinAM.Non-fluorescentCalceinDeepRed™acetatecaneasilygetintolivecellsandhydrolyzestogeneratestronglyfluorescentCalceinDeepRed™(Cat#:21902)dye.CalceinDeepRed™dyecanbemonitoredwiththecommonCy5filterset.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecells:

Plate100to10,000cellsperwellina96-wellblackwall/clearbottommicroplate.Addtestcompoundsintothecellsforadesiredperiodoftime(suchas24,48or96hours)ina37°C,5%CO2incubator.Forblankwells(mediumwithoutthecells),addthecorrespondingamountofcompoundbuffer.Thetotalsuggestedvolumeis100µLfora96-wellplate,and25µLfora384-wellplate.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforproliferationorcytotoxicityinduction.Forproliferation,usefewercells,andfortoxicityassays,usemorecellstostartwith.

 

2.PrepareCalceinDeepRedworkingsolution(for1plate):

2.1   Preparea2to5mMstocksolutionofCalceinDeepRedinhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionshouldbealiquotedandrefrozenat<-20oC.

Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.

 

2.2   PrepareCalceinDeepRedworkingsolution:Onthedayoftheexperiment,eitherdissolveCalceinDeepRedsolidinDMSOorthawanaliquotoftheCalceinDeepRedstocksolutiontoroomtemperature.Preparea5to10µMCalceinDeepRedworkingsolutioninHanksand20mMHepesbuffer(HHBS)orbufferofyourchoicepH7,with2.5mMprobenicid.

Note1:10mLofCalceinDeepRedworkingsolutionisnotstableatroomtemperature;preparefreshbeforeuse.

Note2:Manycellscontainorganic-aniontransporters.Itishighlyrecommendedtoaddadditional2.5mMprobenecid,aninhibitoroforganic-aniontransporters,intheCalceinDeepRedworkingsolution.

 

3.Runthecellviabilityassay:

3.1   Treatcellswithtestcompoundsasdesired(fromStep1).

Note:Itisnotnecessarytowashcellsbeforecompoundaddition.However,iftestedcompoundsareserumsensitive,growthmediumandserumfactorscanbeaspiratedawaybeforeaddingcompounds,providedresidualvolumesaftertheaspiratestep.Alternatively,cellscanbegrowninserum-freemedia.

 

3.2    Removethemediumfromthecells.

Note:MediummustberemovedbeforedyeloADIng.

 

3.3   Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofCalceinDeepRedworkingsolution(fromStep2.2).

 

3.4   IncubatetheCalceinDeepReddye-loadingplateatroomtemperatureor37oCfor1hr,protectedfromlight.

Note1:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.

Note2:Fornon-adherentcells,itisrecommendedtocentrifugecellplatesat800rpmfor2minuteswithbrakeoffaftertheincubationofthedye.

 

3.5   MonitorthefluorescencechangeatEx/Em=635/670nm.

Note:Ifthebackgroundishigh,replacetheCalceinDeepRedworkingsolution(fromStep3.4)withHHBScontaining2.5mMprobenicidbeforemonitoringthefluorescencesignal.

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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