Thermo Fisher Scientific
Live/Dead cell viability/cytotoxicity kit
L3224
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We used this to elucidate the signaling pathway involved in toxicity of natural toxins. During the study to standardize the dose of MAPK pathway inhibitors, we have used this kit to check the viability of IMR-32 cells before and after the treatment with toxin and inhibitors. The kit is easy to optimize and clearly distinguishes live and dead populations.
Fluorescence microscopy
IMR-32 cell line (can be used for any mammalian cell lines)
Culture the IMR-32 cells on coverslips or in chamber slides, at 70–80% confluency treat the cells with your agent and inhibitors. After completion of incubation time, wash the cells twice with PBS and stain the cells with 100–150 µL of dye solution diluted in PBS (20 µL of 2 mM EthD-1 stock solution and 5 µL of 4 mM calcein AM stock solution in 10 mL of D-PBS cell culture grade). Incubate the cells for 30–40 minutes in dark at room temperature and again wash with D-PBS and visualized cells under fluorescence microscope.
Wash the cells properly with D-PBS before staining with dye solution.
Clearly distinguished live and dead cell population in the treated samples. Live cells were visualized as green dots and dead cells as red dots with bright fluorescence.
10.1007/s12035-014-8816-4
None
Easy to optimize and can be used at lesser concentrations mentioned in the protocol.
Excellent Live/dead Viability/cytotoxicity kit to stain the mammalian cells within 30 minutes.