Abstract
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. Its principle is based on the ability of the protein dye sulforhodamine B to bind electrostatically and pH dependent on protein basic amino acid residues of trichloroacetic acid-fixed cells. Under mild acidic conditions it binds to and under mild basic conditions it can be extracted from cells and solubilized for measurement. Results of the SRB assay were linear with cell number and cellular protein measured at cellular densities ranging from 1 to 200‰ of confluence. Its sensitivity is comparable with that of several fluorescence assays and superior to that of Lowry or Bradford. The signal-to-noise ratio is favorable and the resolution is 1000-2000 cells/well. It performed similarly compared to other cytotoxicity assays such as MTT or clonogenic assay. The SRB assay possesses a colorimetric end point and is nondestructive and indefinitely stable. These practical advances make the SRB assay an appropriate and sensitive assay to measure drug-induced cytotoxicity even at large-scale application.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
Rubinstein, L. V., Shoemaker, R. H., Paull, K. D., Simon, R. M., Tosini, S., Skehan, P., Scudiero, D. A., Monks, A., and Boyd, M. R. (1990) Comparison of in vitro anticancer-drug-screening data generated with a tetrazolium assay versus a protein assay against a diverse panel of human tumor cell lines. J. Natl. Cancer Inst. 82, 1113–1118.
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.
Keepers, Y. P., Pizao, P. E., Peters, G. J., van Ark-Otte, J., Winograd, B., and Pinedo, H. M. (1991) Comparison of the sulforhodamine B protein and tetrazolium (MTT) assays for in vitro chemosensitivity testing. Eur. J. Cancer 27, 897–900.
Hodes, L., Paull, K., Koutsoukos, A., and Rubinstein, L. (1992) Exploratory data analytic techniques to evaluate anticancer agents screened in a cell culture panel. J. Biopharm. Stat. 2, 31–48.
Voigt, W., Bulankin, A., Muller, T., Schoeber, C., Grothey, A., Hoang-Vu, C., and Schmoll, H. J. (2000) Schedule-dependent antagonism of gemcitabine and cisplatin in human anaplastic thyroid cancer cell lines. Clin. Cancer Res. 6, 2087–2093.
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.
Berenbaum, M. C. (1989) What is synergy? Pharmacol. Rev. 41, 93–141.
Greco, W. R., Bravo, G., and Parsons, J. C. (1995) The search for synergy: a critical review from a response surface perspective. Pharmacol. Rev. 47, 331–385.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Voigt, W. (2005). Sulforhodamine B Assay and Chemosensitivity. In: Blumenthal, R.D. (eds) Chemosensitivity. Methods in Molecular Medicine™, vol 110. Humana Press. https://doi.org/10.1385/1-59259-869-2:039
Download citation
DOI: https://doi.org/10.1385/1-59259-869-2:039
Publisher Name: Humana Press
Print ISBN: 978-1-58829-345-9
Online ISBN: 978-1-59259-869-4
eBook Packages: Springer Protocols