LiveONLY™ Nuclear Green

1. Prepare the cell samples and treat cells as desired.
2. Add the dye working solution.
3. Incubate for 10 to 30 minutes.
4. Analyze with fluorescence microscope using FITC filter.

Note: Allow dye to warm to room temperature before opening the vials. Because this reagent binds to nucleic acids, it should be treated as a potential mutagen and handled with appropriate care. The DMSO stock solution should be handled with particular caution as DMSO is known to facilitate the entry of organic molecules into tissues.

Add 12 µL of LiveONLY™ Nuclear Green to 10 mL of cell culture medium or HHBS buffer (Cat# 20011) and mix well.

Note: 10 mL volume is sufficient for 100 tests. Make LiveONLY™ Nuclear Green dye working solution sufficient for the assays and use promptly. 

Note: Store unused LiveONLY™ Nuclear Green at -20 °C for further use.

The following protocol can be used as a guideline and can be adapted for any cell type. Growth medium, cell density, the presence of other factors may influence staining.

1. Prepare the cell samples and treat cells as desired in a 96-well plate.
2. Remove the cell culture medium.
3. Add 100 µL of LiveONLY™ Nuclear Green dye working solution and incubate for 10 to 30 minutes at 37°C in a 5% CO₂ incubator, protected from light. (Total volume = 100 µL/well).
   Note: For optimal staining, try a range of dye concentrations to determine the optimal staining.
4. Observe the cells using a fluorescence microscope with FITC filter set.
   Note: Cell culture medium with dye working solution can be removed and be replaced with aqueous buffers such as HHBS buffer, if necessary.