Resazurin sodium

Resazurin sodium is commonly used to measure bacterial and eukaryotic cell viability through its reduction to the fluorescent product resorufin. No viable bacteria are detected 24 h post-inoculation following inclusion of Resazurin sodium in TSBc cultures of F. tularensis LVS at the concentration of 44 μM. Lowering the Resazurin sodium concentration to as little as 4.4 μM still results in a 10-fold reduction in viable F. tularensis LVS compare to growth medium alone. Both Resazurin sodium treatments result in a significant decrease in viable F. tularensis LVS bacteria over 22 h. Treatment with Resazurin sodium significantly reduces the number of viable F. tularensis LVS bacteria in HEK293 cells 22 h post-infection^[1].

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO or ddH2O
Concentration: 1 mg/mL (optimized according to the experiment).
Storage: Store at -20°C or -80°C in dark after aliquoting. Avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute to 1 μg/mL with PBS or serum-free medium (optimized according to the experiment).
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Cell viability
1. Defrost the Resazurin sodium solution in a 37°C water bath.
2. Place the cells on a 96-well plate, and washed by PBS (avoid light).
3. Remove the PBS wash, then add 100 μL (1 μg/mL) of Resazurin sodium solution.
4. Place the plate, in the incubator for 30 min (incubation time depends on cell type and cell number).
5. Measure Resazurin sodium fluorescence (Ex = 530-560 nm, Em = 590 nm) using a microplate reader.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.