Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 646/659 |
MW | ~500 |
CAS# | N/A |
Solvent | DMSO |
Storage | F/D/L |
Category | CellBIOLOGy LabelingCells |
Related | ViABIlityandProliferation CellViability MDRResearch BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
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.PrepareCalceinDeepRed™workingsolution(for1plate):
2.1 Preparea2to5mMstocksolutionofCalceinDeepRed™inhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionshouldbealiquotedandrefrozenat<-20oC.
Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
2.2 PrepareCalceinDeepRed™workingsolution:Onthedayoftheexperiment,eitherdissolveCalceinDeepRed™solidinDMSOorthawanaliquotoftheCalceinDeepRed™stocksolutiontoroomtemperature.Preparea5to10µMCalceinDeepRed™workingsolutioninHanksand20mMHepesbuffer(HHBS)orbufferofyourchoicepH7,with2.5mMprobenicid.
Note1:10mLofCalceinDeepRed™workingsolutionisnotstableatroomtemperature;preparefreshbeforeuse.
Note2:Manycellscontainorganic-aniontransporters.Itishighlyrecommendedtoaddadditional2.5mMprobenecid,aninhibitoroforganic-aniontransporters,intheCalceinDeepRed™workingsolution.
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)ofCalceinDeepRed™workingsolution(fromStep2.2).
3.4 IncubatetheCalceinDeepRed™dye-loadingplateatroomtemperatureor37oCfor1hr,protectedfromlight.
Note1:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
Note2:Fornon-adherentcells,itisrecommendedtocentrifugecellplatesat800rpmfor2minuteswithbrakeoffaftertheincubationofthedye.
3.5 MonitorthefluorescencechangeatEx/Em=635/670nm.
Note:Ifthebackgroundishigh,replacetheCalceinDeepRed™workingsolution(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
AAT Bioquest常用产品报价单
产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |