An improved colorimetric assay for interleukin 2

Hiroko Tada; Osamu Shiho; Ken ichi Kuroshima; Masaru Koyama; Kyozo Tsukamoto

doi:10.1016/0022-1759(86)90183-3

An improved colorimetric assay for interleukin 2

Hiroko Tada, Osamu Shiho, Ken ichi Kuroshima, Masaru Koyama, Kyozo Tsukamoto

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737 Citations (Scopus)

Abstract

Mosmann's method for measuring the number of viable cells with a tetrazolium salt, 3-(4,5-dimethyl-thiazol-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). (1) 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, (2) 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 MIT 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.

Original language	English
Pages (from-to)	157-165
Number of pages	9
Journal	Journal of Immunological Methods
Volume	93
Issue number	2
DOIs	https://doi.org/10.1016/0022-1759(86)90183-3
Publication status	Published - Nov 6 1986
Externally published	Yes

Keywords

Colorimetric assay
Interleukin-2
Tetrazolium salt

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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Tada, H., Shiho, O., Kuroshima, K. I., Koyama, M., & Tsukamoto, K. (1986). An improved colorimetric assay for interleukin 2. Journal of Immunological Methods, 93(2), 157-165. https://doi.org/10.1016/0022-1759(86)90183-3

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abstract = "Mosmann's method for measuring the number of viable cells with a tetrazolium salt, 3-(4,5-dimethyl-thiazol-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). (1) 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, (2) 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 MIT 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.",

keywords = "Colorimetric assay, Interleukin-2, Tetrazolium salt",

author = "Hiroko Tada and Osamu Shiho and Kuroshima, {Ken ichi} and Masaru Koyama and Kyozo Tsukamoto",

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AU - Tada, Hiroko

AU - Shiho, Osamu

AU - Kuroshima, Ken ichi

AU - Koyama, Masaru

AU - Tsukamoto, Kyozo

PY - 1986/11/6

Y1 - 1986/11/6

N2 - Mosmann's method for measuring the number of viable cells with a tetrazolium salt, 3-(4,5-dimethyl-thiazol-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). (1) 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, (2) 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 MIT 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.

AB - Mosmann's method for measuring the number of viable cells with a tetrazolium salt, 3-(4,5-dimethyl-thiazol-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). (1) 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, (2) 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 MIT 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.

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KW - Interleukin-2

KW - Tetrazolium salt

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