Sulforhodamine B colorimetric assay for cytotoxicity screening

Nat Protoc. 2006;1(3):1112-6. doi: 10.1038/nprot.2006.179.


The sulforhodamine B (SRB) assay is used for cell density determination, based on the measurement of cellular protein content. The method described here has been optimized for the toxicity screening of compounds to adherent cells in a 96-well format. After an incubation period, cell monolayers are fixed with 10% (wt/vol) trichloroacetic acid and stained for 30 min, after which the excess dye is removed by washing repeatedly with 1% (vol/vol) acetic acid. The protein-bound dye is dissolved in 10 mM Tris base solution for OD determination at 510 nm using a microplate reader. The results are linear over a 20-fold range of cell numbers and the sensitivity is comparable to those of fluorometric methods. The method not only allows a large number of samples to be tested within a few days, but also requires only simple equipment and inexpensive reagents. The SRB assay is therefore an efficient and highly cost-effective method for screening.

MeSH terms

  • Cell Count / methods*
  • Cell Culture Techniques / methods
  • Colorimetry / methods*
  • Fluorescent Dyes
  • Rhodamines / metabolism*
  • Sensitivity and Specificity
  • Toxicity Tests / methods*


  • Fluorescent Dyes
  • Rhodamines
  • lissamine rhodamine B