An improved colorimetric assay for interleukin 2

J Immunol Methods. 1986 Nov 6;93(2):157-65. doi: 10.1016/0022-1759(86)90183-3.

Abstract

Mosmann's method for measuring the number of viable cells with a tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide (MTT), was modified to make it possible to measure a large number of interleukin 2 (IL-2) samples at one time with less labor and more accuracy. Each step of the method was examined in detail and modified (the modified MTT method). An IL-2-dependent mouse natural killer cell line, NKC3, was used as an indicator cell line. The incubation period before adding MTT was reduced to 24 h, A solution of 10% sodium dodecyl sulfate-0.01 N HCl was used to dissolve the MTT formazan produced. We have compared the values obtained by the modified MTT method and the conventional [3H]thymidine method (3H-TdR method), and confirmed that the estimates of IL-2 content were almost equal. The variation of IL-2 content measured by both methods was within 5% in terms of the standard error.

MeSH terms

  • Cells, Cultured
  • Colorimetry
  • Detergents / pharmacology
  • Humans
  • Interleukin-2 / analysis*
  • Solubility
  • Tetrazolium Salts
  • Thiazoles
  • Tritium / metabolism

Substances

  • Detergents
  • Interleukin-2
  • Tetrazolium Salts
  • Thiazoles
  • Tritium
  • thiazolyl blue