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

Protocol tips

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	Add 2 µM calcein AM and 4 µM EthD-1 Incubate the cells for 30–45 minutes at room temperature

Protocol tips
Add 2 µM calcein AM and 4 µM EthD-1 Incubate the cells for 30–45 minutes at room temperature

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

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells below.

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