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AAT Bioquest/Cell Explorer™ Live Cell Labeling Kit *Orange Fluorescence with 405 nm Excitation*/22616/200 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 398/545 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy LabelingCells |
| Related | FluorescenceImaging BiochemicalAssays |
1.Preparecells:
1.1 Foradherentcells:Platecellsovernightingrowthmediumat10,000to40,000cells/well/100µLfora96-wellplateor2,500to10,000cells/well/25µLfora384-wellplate.
1.2 Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletsinculturemediumat50,000-100,000cells/well/100µLfora96-wellpoly-Dlysineplateor10,000-25,000cells/well/25µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiment.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.
2.PrepareCytoCalcein™Violet550stainsolution:
2.1 PrepareCytoCalcein™Violet550stocksolution:Add20µLofDMSOintoCytoCalcein™Violet550vial(ComponentA),andmixthemwell.
Note:20µLofCytoCalcein™Violet550stocksolutionisenoughfor1plate.UnusedCytoCalcein™Violet550stocksolutioncanbealiquotedandstoredat<-20oCforonemonthifthetubesaresealedtightly.Avoidrepeatedfreeze-thawcyclesandprotectitfromlight.
2.2 PrepareCytoCalcein™Violet550workingsolution:Add20µLofDMSO-reconstitutedCytoCalcein™Violet550stocksolution(fromStep2.1)into10mLofHHBS(ComponentB),andmixthemwell.
3.Stainthecells:
3.1 Removethegrowthmedium.
3.2 Add100µL/well(96-wellplate)or25µL/well(384-wellplate)CytoCalcein™Violet550workingsolution(fromStep2.2)intothecellplate.
3.3 Incubatethecellsina37oC,5%CO2incubatorfor30minutesto1hour.
3.4 RemovetheCytoCalcein™Violet550workingsolutionfromthecells,andwashthecellswithHHBS(ComponentB)for2to3times,andreplacewithHHBS.
3.5 ImagethecellsusingafluorescencemicroscopewithafiltersetatEx/Em=405/550nm.
| References&Citations |  CitationExplorer |
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