TABLE 2:
Confocal Immunofluorescence Imaging Channel Settings
| Channel | Antigen or Target | Antibody Clone | Dye | Ordering Info | Dilution | Excitation (nm) | Emission (nm) | Exposure (ms) | Signal Location |
|---|---|---|---|---|---|---|---|---|---|
| 1 | E-CADHERIN | 36/E-Cadherin | BrilliantViolet-421 | Becton Dickinson, cat. no. 564186 |
1:10 | 405 | 435/40 | 1000 | Surface |
| 2 | NANOG | N31–355 | AlexaFluor-488 | Becton Dickinson cat. no. 560791 |
1:10 | 488 | 510/20 | 1000 | Nuclear |
| 3 | OCT3/4 | 40/Oct-3 | AlexaFluor-555 | Becton Dickinson cat. no. 560306 |
1:10 | 561 | 600/37 | 1000 | Nuclear |
| 4 | TRA-1–60 | TRA-1–60 | AlexaFluor-647 | Becton Dickinson cat. no. 560122 |
1:10 | 640 | 676/29 | 500 | Surface |
| 5 | SSEA-5 | 8e11 | PerCP-Vio700 | Miltenyi, cat. no. 130-106-664 |
1:100 | 488 | 676/29 | 500 | Surface |
| 6 | DNA | n/a | SytoxGreen | ThermoFisher, cat. no. S7020 |
500 nM | 488 | 525/20 | 250 | Nuclear |