Product Pathways - Screening Technologies
Resazurin Cell Viability Kit #11884
| Kit Includes | Quantity |
|---|---|
| Resazurin Solution | 25 ml |
Species Cross-Reactivity
All
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.
Description
The Resazurin Cell Viability Kit is a fluorescent assay that detects cellular metabolic activity. The blue nonfluorescent resazurin reagent is reduced to highly fluorescent resorufin by dehydrogenase enzymes in metabolically active cells. This conversion only occurs in viable cells and thus, the amount of resorufin produced is proportional to the number of viable cells in the sample. The resorufin formed in the assay can be quantified by measuring the relative fluorescence units (RFU) using a fluorometer (Ex=530-570 nm, Em=590-620 nm).
Specificity / Sensitivity
The Resazurin Cell Viability Kit detects resorufin produced from resazurin conversion by metabolic enzymes in live cells. This kit is expected to work in most cell lines. For most experiments, 0.02-2x105 cells/well should be sufficient, but this can vary depending on the cell type and incubation time. For best results, a cell number titration and incubation time course (as shown in Figure 1) is recommended.Note: Microbial contaminants will reduce resazurin to resorufin, yielding false positive results. Autofluorescent compounds may interfere with this assay.
Cell-based assay
Figure 1. HeLa cells were seeded at varying density in a 96-well plate and incubated overnight. The Resazurin solution (10% of cell culture volume) was added to the plate and relative fluorescent units were measured at 0, 1, 2, 4, and 6 hr.
Cell-based assay
Figure 2. Jurkat cells were seeded at 1x105 cells/well in a 96-well plate and then treated overnight with various concentrations of astemizole or terfenadine. Cytotoxicity was measured using the Resazurin Cell Viability Kit (left), followed by BrdU Cell Proliferation Assay Kit #6813 (right).
Background
Cell viability assays are widely used in drug discovery for the study of growth factors, cytokines, and cytotoxic agents. High throughput screening, in both early drug discovery compound screening and subsequent drug safety and toxicity studies, require reliable, sensitive, and simple assays with the ability to analyze a large number of samples. Colorimetric cell viability assays using tetrazolium salt, such as MTT, XTT, and WST-1, were developed based on live cell reduction of tetrazolium salt into highly colored formazan compounds (1,2). Similarly, resazurin (blue and nonfluorescent) can be reduced to resorufin (pink and highly fluorescent) in live cells and is therefore used to assess mammalian cell toxicity, viability, migration, and invasion (3-6). Similar to the XTT Cell Viability Kit #9095, the Resazurin Cell Viability Kit does not require radioactive materials, cell fixation, or cell permeabilization, and cells used in this assay may be used for further analysis.
- Scudiero, D.A. et al. (1988) Cancer Res 48, 4827-33.
- Roehm, N.W. et al. (1991) J Immunol Methods 142, 257-65.
- Nakayama, G.R. et al. (1997) J Immunol Methods 204, 205-8.
- Al-Nasiry, S. et al. (2007) Hum Reprod 22, 1304-9.
- Anoopkumar-Dukie, S. et al. (2005) Br J Radiol 78, 945-7.
- O'Brien, J. et al. (2000) Eur J Biochem 267, 5421-6.
Application References
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Protocols
* Product-specific protocol.
Companion Products
- 9095 XTT Cell Viability Kit
- 6813 BrdU Cell Proliferation Assay Kit
- 5492 BrdU Cell Proliferation Chemiluminescent Assay Kit
For Research Use Only. Not For Use In Diagnostic Procedures.
