An improved colorimetric assay for interleukin 2

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

Research output: Contribution to journalArticle

706 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 languageEnglish
Pages (from-to)157-165
Number of pages9
JournalJournal of Immunological Methods
Volume93
Issue number2
DOIs
Publication statusPublished - Nov 6 1986
Externally publishedYes

Fingerprint

Interleukin-2
Tetrazolium Salts
Cell Line
Bromides
Sodium Dodecyl Sulfate
Natural Killer Cells
Thymidine
Cell Count

Keywords

  • Colorimetric assay
  • Interleukin-2
  • Tetrazolium salt

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

An improved colorimetric assay for interleukin 2. / Tada, Hiroko; Shiho, Osamu; Kuroshima, Ken ichi; Koyama, Masaru; Tsukamoto, Kyozo.

In: Journal of Immunological Methods, Vol. 93, No. 2, 06.11.1986, p. 157-165.

Research output: Contribution to journalArticle

Tada, H, Shiho, O, Kuroshima, KI, Koyama, M & Tsukamoto, K 1986, 'An improved colorimetric assay for interleukin 2', Journal of Immunological Methods, vol. 93, no. 2, pp. 157-165. https://doi.org/10.1016/0022-1759(86)90183-3
Tada, Hiroko ; Shiho, Osamu ; Kuroshima, Ken ichi ; Koyama, Masaru ; Tsukamoto, Kyozo. / An improved colorimetric assay for interleukin 2. In: Journal of Immunological Methods. 1986 ; Vol. 93, No. 2. pp. 157-165.
@article{3750e674fb5a4a5e8f631328d26d37a0,
title = "An improved colorimetric assay for interleukin 2",
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",
year = "1986",
month = "11",
day = "6",
doi = "10.1016/0022-1759(86)90183-3",
language = "English",
volume = "93",
pages = "157--165",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - An improved colorimetric assay for interleukin 2

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.

KW - Colorimetric assay

KW - Interleukin-2

KW - Tetrazolium salt

UR - http://www.scopus.com/inward/record.url?scp=0023022289&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023022289&partnerID=8YFLogxK

U2 - 10.1016/0022-1759(86)90183-3

DO - 10.1016/0022-1759(86)90183-3

M3 - Article

VL - 93

SP - 157

EP - 165

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 2

ER -