Cytotoxicity Detection Kit (LDH)

Protocol tips

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	Incubate the cells in an incubator (37°C, 5% CO2, 90% humidity)

Protocol tips
Incubate the cells in an incubator (37°C, 5% CO2, 90% humidity)

Publication protocol

The safety of B. subtilis (natto) B4 spores was determined by cytotoxicity evaluation, which included a cell viability test and LDH leakage assay. The viability of RAW 264.7 cells was measured by trypan blue dye exclusion test as described by Sarir et al. (23) and calculated according to the following formula: cell viability (%) = NV× 100/NT where NV is the number of viable cells and NT the total number of cells. The amount of LDH leakage from necrotic macrophages into the culture supernatant was determined by the method described by Weyermann et al. (24) using a Cytotoxicity Detection Kit (LDH) (Roche Diagnostics, Mannheim, Germany).

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Manufacturer protocol

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