Content #1
Content #1
Content #1
Resazurin (also known as alamarBlue™) is the N-oxide of the fluorescent dye resorufin and is useful for detecting reductive activities in cells. It has been widely used for measuring cell proliferation (1,2) and mitochondrial metabolic activity. Resazurin itself is non-fluorescent until it is reduced to the highly red fluorescent resorufin (λEx/λEm: 563/587 nm).
Resazurin (also known as alamarBlue™) is the N-oxide of the fluorescent dye resorufin and is useful for detecting reductive activities in cells. It has been widely used for measuring cell proliferation (1,2) and mitochondrial metabolic activity. Resazurin itself is non-fluorescent until it is reduced to the highly red fluorescent resorufin (λEx/λEm: 563/587 nm). Usually, NADPH or NADH is the reductant that converts resazurin to resorufin in the presence of diaphorase as the enzyme. Thus, resazurin can be used to detect NADH, NADPH, or diaphorase levels. Furthermore, the resazurin/diaphorase/NADPH system can be used to detect any biochemical or enzyme activity that is involved in a biochemical reaction generating NADH or NADPH (3-9). Although resazurin is available from most of the chemical suppliers, a trace amount of resorufin contaminant often makes the material unsuitable for bioassay applications. We supply a high grade resazurin that has a minimal background fluorescence. Please also see our ready-to-use Resazurin Cell Viability Assay Kit (30025).
1. J Ocular Pharmacol Therapeut 26, 1 (2010).
2. J Clin Lab Anal 9, 89 (1995).
3. J Immunol Methods 175, 181 (1994).
4. J Anal Toxicol 8, 273 (1984).
5. Clin Chem 29, 171 (1983).
6. Clin Chim Acta 107, 149 (1980).
7. Clin Chem 26, 61 (1980).
8. Biochem Biophys Acta 484, 249 (1977).
1. J Ocular Pharmacol Therapeut 26, 1 (2010).
2. J Clin Lab Anal 9, 89 (1995).
3. J Immunol Methods 175, 181 (1994).
4. J Anal Toxicol 8, 273 (1984).
5. Clin Chem 29, 171 (1983).
6. Clin Chim Acta 107, 149 (1980).
7. Clin Chem 26, 61 (1980).
8. Biochem Biophys Acta 484, 249 (1977).
Medium with phenol red is compatible with the resazurin assay. Phenol red does not interfere with the resazurin reaction nor does it affect detection.
AlamarBlue® contains resazurin and additional compounds to prevent the over-reduction of resazurin to a non-fluorescent product. These additives also slow the rate of generation of the fluorescent product. Consequently, the alamarBlue® assay requires longer incubation times compared to resazurin.
Resazurin is reduced by cells to the fluorescent product resorufin. Resorufin can be reduced further to a non-fluorescent compound. Therefore the densest wells may have lowest fluorescence due to over-reduction of the substrate. Please see the product information sheet for more details. The kit protocol provides general guidelines and may need to be optimized empirically for your experimental system. You may need to vary cell density or assay incubation time to ensure that your samples fall in the linear range of the kit.