| Field | Specification |
|---|---|
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Cell culture |
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Overview
Homogeneous assay for cell viability, proliferation, cytotoxcity and high-throughput screen for anticancer agents. The assay uses FL530/590nm for signal readout. Compatible sample input includes Cell culture. Typical stated assay timing is Assay takes 1-5 hrs, hands-on time 30 min.
Key elements and design rationale
- Readout format: FL530/590nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Cell culture, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 100 cells for interpreting low-signal samples.
- Feature emphasis: Safe. Non-radioactive assay (cf. 3 H-thymidine incorporation assay).
Additional feature notes highlight Sensitive and accurate. As low as 100 cells can be accurately quantified; Time efficient. High-throughput assay in 96-well and 384-well plates allows the simultaneous processing of tens of thousands of samples per day. Available format information for this listing includes 5000 Tests.
Biological background
This product is centered on measurement of cell viability within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
This homogeneous assay involves simply adding a single reagent, the CellQuanti-Blue™ reagent, to the cell culture and measuring the fluorescence intensity (excitation wavelength = 530 – 570 nm, emission wavelength = 590 – 620 nm) after an incubation step. The CellQuanti-Blue™ reagent, like other resazurin-based assays such as the Alamar Blue reagent, utilizes the redox dye resazurin which is not fluorescent, but upon reduction by metabolically active cells is converted into a highly fluorescent product (resorufin). Living cells can readily reduce this non-toxic reagent and the resulting increase in fluorescence intensity can be conveniently monitored using a fluorescence spectrophotometer or plate reader. Nonviable cells have no metabolic capacity and, thus, will not reduce the dye. Therefore, the fluorescence intensity observed in this assay is a true measure of the viable cells. The CellQuanti-Blue™ reagent has been optimized for maximum sensitivity, reproducibility and long shelf-life. The homogeneous cell-based assay can be performed in multi-well plates. The reagent is compatible with all culture media and with all liquid handling systems for high-throughput screening applications in 96-well and 384-well plates. Applications include cell proliferation, cytotoxicity and apoptosis.
Detection method
Fluorescent (FL 530/590 nm).
Detection limit and analytical sensitivity
Reported detection limit: 100 cells.
Procedures and timing
Stated procedure or timing information: Assay takes 1-5 hrs, hands-on time 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify cell viability or cytotoxicity in cultured cells by FL530/590 nm readout.
- Compare dose-response effects of compounds across treated cell-culture wells.
- Monitor cytotoxicity time-courses after toxin, detergent, or drug exposure in culture.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
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 FITC-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 laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Olaparib potentiates anticancer drug cytotoxicity via 53bp1 in oesophageal squamous cell carcinoma cells
Miyamoto, K., et al (2020). Olaparib potentiates anticancer drug cytotoxicity via 53bp1 in oesophageal squamous cell carcinoma cells. Anticancer Research, 40(2), 813-823. Assay: Cell Viability in human esophogeal carcinoma cells.
Swelling characteristics and biocompatibility of ionic liquid based hydrogels for biomedical applications
Claus, J.,et al. (2020). Swelling characteristics and biocompatibility of ionic liquid based hydrogels for biomedical applications. PLOS ONE, 15(4), e0231421 Assay: Cell Viability in mouse fibroblast cells.
Potent double prodrug forms of synthetic phosphoantigens
Harmon, N. M.,et al. (2020). Potent double prodrug forms of synthetic phosphoantigens. Bioorganic & Medicinal Chemistry, 28(19), 115666. Assay: Cell Viability in human PBMC cells.
Methods of reducing levels of tau
Kim, TW et al (2019). Methods of reducing levels of tau. US Patent 9,696,306. Assay: Cell viability in human cells.
Synergistic Effects of Olaparib and DNA-damaging Agents in Oesophageal Squamous Cell Carcinoma Cell Lines
Miyamoto K et al (2019). Synergistic Effects of Olaparib and DNA-damaging Agents in Oesophageal Squamous Cell Carcinoma Cell Lines. Anticancer Res. 39(4):1813-1820. Assay: Cell viability in human cell lines.
Therapeutic and diagnostic methods for autism spectrum disorders and other conditions
Nath, A et al (2019). Therapeutic and diagnostic methods for autism spectrum disorders and other conditions. US20190025327A1 Assay: Cell viability in rat fetal cells.
CD147 As A Potential Therapeutic Target In Glioblastoma Treatment” All Nmu Master’S Theses
Adams, Beau (2018). CD147 As A Potential Therapeutic Target In Glioblastoma Treatment” All Nmu Master’S Theses. 565. Assay: Cell viability in mice glioblastoma cells.
Activated T cells induce proliferation of oligodendrocyte progenitor cells via release of vascular endothelial cell growth factor-A
Choi EH et al (2018). Activated T cells induce proliferation of oligodendrocyte progenitor cells via release of vascular endothelial cell growth factor-A. Glia.66(11):2503-2513. Assay: Cell viability in human hematopoietic stem cells.
Augmentation of the cytotoxic effects of nitrogen-containing bisphosphonates in hypoxia
Minegaki, T (2018). Augmentation of the cytotoxic effects of nitrogen-containing bisphosphonates in hypoxia. J Pharm Pharmacol. 70(8):1040-1047. Assay: Cell viability in human cancer cells.
Histone deacetylase inhibitors sensitize 5-fluorouracil-resistant MDA-MB-468 breast cancer cells to 5-fluorouracil
Minegaki, T et al (2018). Histone deacetylase inhibitors sensitize 5-fluorouracil-resistant MDA-MB-468 breast cancer cells to 5-fluorouracil. Oncol Lett. 16(5):6202-6208. Assay: Cell viability in human breast cancer cell line.
Evaluation of Trace Elements in Augmentation of Statin-Induced Cytotoxicity in Uremic Serum-Exposed human Rhabdomyosarcoma Cells
Uchiyama H et al (2018). Evaluation of Trace Elements in Augmentation of Statin-Induced Cytotoxicity in Uremic Serum-Exposed human Rhabdomyosarcoma Cells. Toxins (Basel). 10(2). pii: E53. Assay: Cell viability in human cells.
Novel approach for a PTX/VEGF dual drug delivery system in cardiovascular applications-an innovative bulk and surface drug immobilization
Wulf K (2018). Novel approach for a PTX/VEGF dual drug delivery system in cardiovascular applications-an innovative bulk and surface drug immobilization. Drug Deliv Transl Res. 8(3):719-728. Assay: Cell viability in human epithelial cells.
Preventive Activity of ar-Turmerone and Bisacurone Isolated from Turmeric Extract Against Ethanol-induced Hepatocyte Injury
Megumi C et al (2017). Preventive Activity of ar-Turmerone and Bisacurone Isolated from Turmeric Extract Against Ethanol-induced Hepatocyte Injury. Food Science and Technology Research, 23 (2), 275-281. Assay: Cell viability in mice hepatocytes.
Layer-by-layer nanoparticle platform for cancer active targeting
Suh MS et al (2017). Layer-by-layer nanoparticle platform for cancer active targeting. Int J Pharm. 517(1-2):58-66. Assay: Cell viability in human lung cancer cells.
Inhibitory Effects of Juices Prepared from Individual Vegetables on CYP3A4 Activity in Recombinant CYP3A4 and LS180 Cells
Tsujimoto M et al (2017). Inhibitory Effects of Juices Prepared from Individual Vegetables on CYP3A4 Activity in Recombinant CYP3A4 and LS180 Cells. Biol Pharm Bull. 40(9):1561-1565. Assay: Cell viability in human cell lines.
Butyrate modifies intestinal barrier function in IPEC-J2 cells through a selective upregulation of tight junction proteins and activation of the Akt signaling pathway
Yan H, Ajuwon KM (2017). Butyrate modifies intestinal barrier function in IPEC-J2 cells through a selective upregulation of tight junction proteins and activation of the Akt signaling pathway. PLoS One. 12(6):e0179586. Assay: Cell viability in pig epithelial cells.
Cytotoxicity of various chemicals and mycotoxins in fresh primary duck embryonic fibroblasts: a comparison to HepG2 cells
Chen, X. et al (2016). Cytotoxicity of various chemicals and mycotoxins in fresh primary duck embryonic fibroblasts: a comparison to HepG2 cells. J Appl Toxicol. 36(11):1437-45. Assay: Cell viability in duck embryonic fibroblasts.
Aqueous extract of Corchorus olitorius decreases cytotoxicity of aflatoxin B atoxin B1and fumonisin B and fumonisin B1 in H4IIE- in H4IIE-luccells
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.
Examination of Cell Viability using the primary culture system of skin fibroblasts
Fujimoto, M et al (2011). Examination of Cell Viability using the primary culture system of skin fibroblasts. Assay: Cell viability in plant fibroblasts.
Examination of Cell Viability using the primary culture system of skin fibroblasts
Fujimoto, M et al (2011). Examination of Cell Viability using the primary culture system of skin fibroblasts. Cell Viabilityof skin fibroblast 2: 23-26. Assay: Cell viability in human fibroblasts.
Methyl Vitamin B12 but not methylfolate rescues a motor neuron-like cell line from homocysteine-mediated cell death
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.
Therapeutic window for bioactive nanocomposites fabricated by laser ablation in polymer-doped organic liquids
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.
YPEL3, a p53-regulated gene that induces cellular senescence
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.
Effect of oxidative stress on splicing of parkin gene
Kuroda, Y et al (2010). Effect of oxidative stress on splicing of parkin gene. Assay: Cell viability in human cell lines.
Effects of weak static magnetic fields on endothelial 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.
Maintenance of paracellular barrier function by insulin-like growth factor-I in submandibular gland 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.
Cell viability assessment: toward content-rich platforms
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.
Activated T-cells inhibit neurogenesis by releasing granzyme B: rescue by Kv1
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.
Alterations in gene expression and sensitivity to genotoxic stress following HdmX or Hdm2 knockdown in human tumor cells harboring wild-type p53
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.
Neuroprotective effects of Brazilian green propolis and its main constituents against oxygen-glucose deprivation stress, with a gene-expression analysis
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.
Preservation of intact adult rat photoreceptors in vitro: study of dissociation techniques and the effect of light
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.
Functional roles for the striatal-enriched transcription factor, Bcl11b, in the control of striatal gene expression and transcriptional dysregulation in Huntington’s disease
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.
Huperzine A provides neuroprotection against several cell death inducers using in vitro model systems of motor neuron cell death
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 cell lines.
Chloroquine mediated molecular tuning of astrocytes for enhanced permissiveness to HIV infection
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.
Geranylgeranylacetone Ameliorates Inflammatory Response to Lipopolysaccharide (LPS) in Murine Macrophages: Inhibition of LPS Binding to The Cell Surface
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(2):115-23. Assay: Cell viability in mouse cell lines.
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