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AAT Bioquest/JC-1 [5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide] *CAS#: 3520-43-2*/22200/5 mg
![AAT Bioquest/JC-1 [5,5,6,6-Tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide] *CAS#: 3520-43-2*/22200/5 mg](https://www.ebiomall.cn/images/AAT201712/chemical-structure-for-jc-1-5-5-6-6-tetrachloro-1-1-3-3-tetraethylbenzimidazolylcarbocyanine-iodide-cas-3520-43-2.png)
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 515/529 |
| MW | 652.23 |
| CAS# | 3520-43-2 |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | CellAnalysis CellApoptosis |
| Related | VitalStains NeurologicalStains BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.PrepareJC-1workingsolution:
1.1 Preparea2to10mMstocksolutionofJC-1inhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionshouldbealiquotedandfrozenat<-20oC.
Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
1.2 Preparea1XJC-1workingsolution:Onthedayoftheexperiment,eitherdissolveJC-1solidinDMSOorthawanaliquotoftheJC-1stocksolutiontoroomtemperature.Preparea10to30µM1XJC-1workingsolutioninHanksand20mMHepesbuffer(HHBS)orbufferofyourchoice,pH7with0.02%Pluronic®F-127.Mixthemwellbyvotexing.
Note:JC-1isnotwatersoluble,soitintendstoaggregateinsolution.ItisrecommendedtofiltertheJC-1workingsolutionbeforeloADIngitintothecells.
2.RunJC-1assaywithafluorescencemicroplatereader:
2.1 Treatcellswithtestcompoundsforadesiredperiodoftime(Forexample,Jurkatcellscanbetreatedwithcamptothecinfor4-6hours)toinduceapoptosis.Forblankwells(mediumwithoutthecells),addthecorrespondingamountofcompoundbuffer.
2.2 Add100µL/well/96-wellplateor25µL/well/384-wellplateofJC-1workingsolution(fromStep1.2)intothecellplate.
2.3 IncubatetheJC-1loadingplateina37oC,5%CO2incubatorfor15-60min.
Note:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
2.4 RemovetheJC-1workingsolutionfromtheplate,washthecellswithHHBSorbufferofyourchoice.Add100µL/well/96-wellplateor25µL/well/384-wellplateofHHBSbacktothecellplate.
2.5 MonitorthefluorescencechangeatEx/Em=490/525nmand490/590nmforratioanalysis.
3.RunJC-1assaywithafluorescencemicroscopeoraflowcytometer:
3.1 Treatcellswithtestcompoundsforadesiredperiodoftime(Forexample,Jurkatcellscanbetreatedwithcamptothecinfor4-6hours)toinduceapoptosis.
3.2 Centrifugethecellstoget1-5x105cellspertube.
3.3 ResUSPendcellsin500µLofJC-1workingsolution(fromStep1.2).
3.4 Incubateatroomtemperatureor37°Cfor10to30min,protectedfromlight.
3.5 WashthecellswithHHBSorbufferofyourchoice.Resuspendcellsin500µLofHHBStoget1-5x105cellspertube.
3.6 MonitorthefluorescencechangeatEx/Em=490/525nmand490/590nmwithafluorescencemicroscope(usingFITCandTRITCfilters)oraflowcytometer(usingFL1andFL2channels).
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