The sulforhodamine B (SRB) assay was developed by Skehan and colleagues to measure drug-induced cytotoxicity and cell proliferation for large-scale drug-screening applications. Sulforhodamine B, an anionic aminoxanthene dye, can form an electrostatic complex with the basic amino acid residues of proteins under moderately acid conditions, which provides a sensitive linear response with cell number and cellular protein measured at cellular densities ranging from 1 to 200% of confluence. The SRB assay possesses a nondestructive and indefinitely stable colorimetric end point and the color development is rapid and stable and is readily measured at absorbance between 560 and 580nm. With many practical advances including a favorable signal-to-noise ratio and a resolution of 1000–2000 cells/well, the SRB assay serves as an appropriate and sensitive assay to measure drug-induced cytotoxicity even at large-scale application.
Creative Bioarray Advantages
- High sensitivity
- Accurate and reproducible
- Low signal-to-noise ratio
- High resolution (1000–2000 cells/well)
- Linear results ranging from 1 to 200% of confluence
- colorimetric, nondestructive, and indefinitely stable end point
- Rapid and low-cost
- Suitable for high-throughput screening
- Seeding of Microtiter Plates for Growth Kinetics
- Choice of Drug Dose Range and Seeding of Culture Plates for Cytotoxicity Assays
- SRB Procedure
- Data Processing and Interpretation of Results
- Cell growth/proliferation
- Cytotoxicity measurement/screening
Figure 1. Graphic determination of IC50
Figure 2. Effects of compounds on cell proliferation
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- Skehan, P., Storeng, R., Scudiero, D., Monks, A., McMahon, J., Vistica, D., Warren, J. T., Bokesch, H., Kenney, S., and Boyd, M. R. (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J. Natl. Cancer Inst. 82, 1107–1112.
- Perez, R. P., Godwin, A. K., Handel, L. M., and Hamilton, T. C. (1993) A comparison of clonogenic, microtetrazolium and sulforhodamine B assays for determination of cisplatin cytotoxicity in human ovarian carcinoma cell lines. Eur. J. Cancer 29A, 395–399.
- Griffon, G., Merlin, J. L., and Marchal, C. (1995) Comparison of sulforhodamine B, tetrazolium and clonogenic assays for in vitro radiosensitivity testing in human ovarian cell lines. Anticancer Drugs 6, 115–123.
- Papazisis, K. T., Geromichalos, G. D., Dimitriadis, K. A., and Kortsaris, A. H. (1997) Optimization of the sulforhodamine B colorimetric assay. J. Immunol. Methods 208, 151–158.
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