Live and Dead Cell Assay (Abcam)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 4 μM of calcein acetomethoxy (calcein AM-5 μL) and 2 mM of ethidium-bromide. Incubate cells for 30-40 min at room temperature in the dark.	It is recommended to analyze the cells as quickly after staining as possible. The staining solution should not be exchanged for another buffer before data acquisition. Stained cells are stable for at least one hour at room temperature after staining. Live and Dead dye staining is lost if cells are fixed.

Protocol tips
Add 4 μM of calcein acetomethoxy (calcein AM-5 μL) and 2 mM of ethidium-bromide. Incubate cells for 30-40 min at room temperature in the dark.
Downstream tips
It is recommended to analyze the cells as quickly after staining as possible. The staining solution should not be exchanged for another buffer before data acquisition. Stained cells are stable for at least one hour at room temperature after staining. Live and Dead dye staining is lost if cells are fixed.

Publication protocol

Cell-drug interaction was also evaluated using cell viability assay (live-dead staining kit) over the period of 21 days. From the stock solution, 4 μM of calcein-acetomethoxy (calcein AM-5 μL) and 2 mM of ethidium-bromide (20 μL) were diluted in 10 mL PBS (tissue culture grade). 20–50 μL of working solution was added to each well and plates were incubated for 30–45 min and labeled cells were observed under Nikon fluorescent microscope as per general protocol [23].

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