Revision 2

#11884Store at -20C

Cell Signaling Technology

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Product Information

Storage

Store at -20°C. Note: Resazurin solution should be stored at -20°C in the dark for long term storage. It can be stored at 4°C in the dark for up to 24 months.

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.

Species Reactivity:

All Species Expected

Source / Purification

Resazurin sodium salt was purched from Sigma, Cat#R7017-5G, Lot#MKBF7963V

Product 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).

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.

  1. Scudiero, D.A. et al. (1988) Cancer Res 48, 4827-33.
  2. Roehm, N.W. et al. (1991) J Immunol Methods 142, 257-65.
  3. Nakayama, G.R. et al. (1997) J Immunol Methods 204, 205-8.
  4. Al-Nasiry, S. et al. (2007) Hum Reprod 22, 1304-9.
  5. Anoopkumar-Dukie, S. et al. (2005) Br J Radiol 78, 945-7.
  6. O'Brien, J. et al. (2000) Eur J Biochem 267, 5421-6.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 2
#11884

Resazurin Cell Viability Kit

Resazurin Cell Viability Kit: Image 1 Expand Image
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.
Resazurin Cell Viability Kit: Image 2 Expand Image
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).