LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy

Protocol tips

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	Mix 35 µL of Component A with 35 µL of Component B in a microfuge tube. Add 6 µL of the combined reagent mixture to te tubes. Incubate at room temperature in the dark for 15 minutes

Protocol tips
Mix 35 µL of Component A with 35 µL of Component B in a microfuge tube. Add 6 µL of the combined reagent mixture to te tubes. Incubate at room temperature in the dark for 15 minutes

Publication protocol

Infected M1-MDMs with knockdown of Rev-erbα or ectopically overexpressed Rev-erbα were lysed with 0.06% SDS after 24 h of incubation, and bacterial suspensions were used with the LIVE/DEAD BacLight bacterial viability and counting kit as per the manufacturer's instructions (Invitrogen). The percentage of live and dead bacteria was determined by flow cytometry (BD FACSCalibur, BD Biosciences) after staining with SYTO9 and propidium iodide. For CFU determination, after macrophage solubilization, the bacterial suspensions were serially diluted, 50 μl of each sample was plated, and CFU were counted. Final calculations included the dilution factor and the volume of diluted sample for plating. Anti-IL10 neutralizing antibody (purified Rat anti-human IL10, BD Biosciences) or isotype control was used 30 min prior to infection with H37Ra or H37Rv.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy below.

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