Membrane integrity–based viability assay
NucRed® Dead 647 ReadyProbes® Reagent is a cell-impermeant stain that emits bright far-red fluorescence when bound to DNA. Since this reagent stains cells that lack plasma membrane integrity, it is extremely useful to measure viability and to measure cytotoxicity in kinetic live-cell assays. NucRed® Dead 647 reagent is also suitable for staining nuclei in fixed-cell preparations and tissue sections.
This protocol can be used for:
- Identifying dead cells using a fluorescence microscope
This protocol should not be used for:
- Flow cytometry
1. Culture cells in appropriate medium and vessel for microscopy
2. Add 2 drops NucRed® Dead 647 reagent per milliliter of medium to label cells
3. Incubate 5–15 minutes
4. Image cells
Spectral information and storage
|NucRed® Dead 647 ReadyProbes® Reagent|
|Standard filter set||Cy®5|
|EVOS® Light Cube||Cy®5|
|Storage conditions||Room temperature|
Tubulin and nuclear staining in HeLa cells. HeLa cells were fixed and permeabilized and stained for tubulin (biotinylated mouse anti-a-tubulin mouse monoclonal antibody followed by an Alexa Fluor 488 goat anti-mouse secondary antibody and counter-stained with NucRed Dead 647 ReadyProbes reagent.