CellQuanti-Blue™ Cell Viability Assay Kit

Which enzymatic reaction converts the resazurin into the fluoresence dye?

The exact mechanism how resazurin is converted into the fluorescent resorufin is not known, but it is generally believed that resazurin enters the cell and is reduced by enzymes in the mitochondrial respiratory chain. As only living cells harbor active mitochondria, the conversion of this dye as measured by the fluorescence intensity of the product, is an accurate measure of mitochondria activity and hence living cells.

I have access to two plate readers: Molecular Devices Flexstation and Victor 3 Perkin Elmer. Do you have info on the optimal excitation and emission wavelength on these specific devices?

Any excitation filter in the range of 530-570 nm combined with a suitable emission filter in the range of 585-620 nm for emission, will work. Because excitation and emission wavelengths are close together, we recommend the use of a cut-off filter, if available.

I am going to do the assay directly on the wells containing media and adherent cells. After the assay, I am interested to harvest the cells for IFA testing (recover of cells with trypsin digestion, then incubate with antibodies, then with FTIC-antibodies, finally viewing under the microscope. Does the dye in CQBL absorbed by the cells? Will it interfere with the IFA viewing under the microscope?

Resazurin enters the cells and it is in the cells that it is reduced into the fluorescent dye. During the wash steps in the antibody staining procedure it will likely be completely removed from the cells. Even it that was not the case, FITC is typically measured at an emission wavelength of 525 nm, and the reduced form of resazurin does not fluoresce at wavelengths below 530 nm. So the assay should not interfere with the immunofluorescence staining.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.

Ibrihim, MIM et al (2014). Aqueous extract of Corchorus olitorius decreases cytotoxicity of aflatoxin B atoxin B1and fumonisin B and fumonisin B1 in H4IIE- in H4IIE-luccells.Hepatoma Research. 1(2) Assay: Cell viability in rat hepatoma cells.Fujimoto, M et al (2011). Examination of Cell Viability using the primary culture system of skin fibroblasts. XXXXX. Assay: Cell viability in human fibroblasts.Hemendinger, RA et al (2011). Methyl Vitamin B12 but not methylfolate rescues a motor neuron-like cell line from homocysteine-mediated cell death. Toxicol Appl Pharmacol. 251(3):217-25. Assay: Cell viability in human cell lines.Hahn, A et al (2010). Therapeutic window for bioactive nanocomposites fabricated by laser ablation in polymer-doped organic liquids. Adv. Eng. Mater. 12(5). Assay: Cell viability in mouse cell lines.Kelley, KD et al (2010). YPEL3, a p53-regulated gene that induces cellular senescence. Cancer Res. 70(9):3566-75. Assay: Cell viability in human U2OS-TetR cells.Martino, CF et al (2010). Effects of weak static magnetic fields on endothelial cells. Bioelectromagnetics 31(4):296-301. Assay: Cell viability in human endothelial cells.Mitsui, R et al (2010). Maintenance of paracellular barrier function by insulin-like growth factor-I in submandibular gland cells. Arch Oral Biol. 55(12):963-9. Assay: Cell viability in rat cells.Ramirez, CN et al (2010). Cell viability assessment: toward content-rich platforms. Expert Opinion on Drug Discovery 5(3): 223-233. Assay: Cell viability in human cell line.Wang, T et al (2010). Activated T-cells inhibit neurogenesis by releasing granzyme B: rescue by Kv1.3 blockers. J Neurosci. 30(14):5020-7. Assay: Cell viability in human neural progenitor cells.Heminger, K et al (2009). Alterations in gene expression and sensitivity to genotoxic stress following HdmX or Hdm2 knockdown in human tumor cells harboring wild-type p53. Aging 1(1):89-108. Assay: Cell viability in human cell lines.Nakajima, Y et al (2009). Neuroprotective effects of Brazilian green propolis and its main constituents against oxygen-glucose deprivation stress, with a gene-expression analysis. Phytother Res. 23(10):1431-8. Assay: Cell viability in rat cell line.Zayas-Santiago A, Kang Derwent JJ (2009). Preservation of intact adult rat photoreceptors in vitro: study of dissociation techniques and the effect of light. Mol Vis.15:1-9. Assay: Cell viability in rat photoreceptor cells.Desplats, PA et al (2008). Functional roles for the striatal-enriched transcription factor, Bcl11b, in the control of striatal gene expression and transcriptional dysregulation in Huntington"s disease. Neurobiol Dis. 31(3):298-308. Assay: Cell viability in human cells.Hemendinger, RA et al (2008). Huperzine A provides neuroprotection against several cell death inducers using in vitro model systems of motor neuron cell death. Neurotox Res. 13(1):49-61. Assay: Cell viability in rat cells.Vijaykumar, TS et al (2008). Chloroquine mediated molecular tuning of astrocytes for enhanced permissiveness to HIV infection. Virol. 381(1):1-5. Assay: Cell viability in human cells.Mochida, S et al (2007). Geranylgeranylacetone Ameliorates Inflammatory Response to Lipopolysaccharide (LPS) in Murine Macrophages: Inhibition of LPS Binding to The Cell Surface. J. Clin. Biochem. Nutr., 41, 115-123. Assay: Cell viability in mouse cell lines. To find more recent publications, pleaseclick here.