Resazurin can be used quantitatively determine cell proliferation, viability, and cytotoxicity. Resazurin, when added to viable cells, is reduced by the cellular enzymatic or chemical reactions converting blue/non-fluorescent resazurin to highly fluorescent resorufin. The assay is simple to perform since the indicator is water-soluble and has low toxicity, thus eliminating the washing/fixing and extraction steps required in other commonly used cell proliferation assays.
Direction for Use
Resazurin may be added at any time point during the culture period. For measurement of cell proliferation, it is best to add resazurin during the log phase of growth.
1. In a sterile work area, add resazurin ready-to-use solution at a volume equal to 10% of the cell culture media volume.
2. Return cells to the incubator and continue the incubation for 2-24 hours*. *Incubation times may vary depending on the metabolic rates of the cell lines being tested.
3. Measure the fluorescent intensity using 540-570 nm excitation and 590 nm emission wavelengths. Absorbance can be measured using a spectrophotometer set at 570 nm.
Storage
Upon receipt the product should be stored at 4 °C, protected from light, and used within 6 months. Avoid repeated freezing and thawing.
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