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11884
Resazurin Cell Viability Kit
Cellular Assay Kits
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Resazurin Cell Viability Kit #11884

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Resazurin Cell Viability Kit: Image 1

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

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).

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 12 months.

Protocol

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Resazurin Cell Viability Kit Protocol

  1. Thaw out Resazurin solution (if kept frozen) and warm it to 37°C to ensure all components are completely in solution.
  2. Plate cells in 96-well plate (black plate with clear bottom). Typical seed cell number is 0.02–2 x 105 cells/well depending on cell growth rate. Cell number titration is recommended to determine the optimal cell seeding density.
  3. Incubate cells with compound of interest for desired period of time (1–72 hr). Make sure all the wells contain the same volume of medium.
  4. Add Resazurin solution to plate (10% of the initial volume in the well). For example, for plates containing 100 μl medium/well, add 10 μl resazurin solution to each well.
  5. Incubate the plate for 1–6 hr in standard culture conditions. Incubation time depends on cell type and cell number. The plate can be read multiple times to determine the optimal time point.
  6. Measure the relative fluorescent units (RFU) using a plate reader. Ex = 530–570 nm, Em = 590–620 nm.

posted February 1, 2012

Protocol Id: 80

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.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
To Purchase # 11884S
Product # Size Price
11884S
25 ml each  (2500 assays (96 well format)) $ 176