LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells

Protocol tips

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	Add ethidium-calcein mixture, and incubate for 30-45 min at RT. A shorter incubation time may be used if the dye concentrations or incubation temperature are increased.

Protocol tips
Add ethidium-calcein mixture, and incubate for 30-45 min at RT. A shorter incubation time may be used if the dye concentrations or incubation temperature are increased.

Publication protocol

Neonatal cardiomyocytes, ESCs, and noncardiomyocytes including MEFs, HepG2, HEK293, peripheral lymphatic cells, C2C12, skeletal myotubes, and fetal neurons were exposed to glucose-free DMEM (Invitrogen) supplemented with or without lactate (Wako). Cell viabilities were determined by the LIVE/DEAD Viability/Cytotoxicity Assay Kit (Invitrogen) based on the simultaneous determination of live and dead cells with the calcein AM and ethidium homodimer-1 probes. Fluorescence imaging of the cells (live cells were labeled green, whereas the nuclei of dead cells were labeled red) was performed with fluorescence microscopy (IX70 microscope; Olympus) equipped with a color charge-coupled device camera (CS220; Olympus). The green-labeled live area was measured using Image J. Relative cell viabilities were calculated in percentages, compared with those before treatment.

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Manufacturer protocol

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