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 | |
| Publication status | Published - Nov 6 1986 |
| Externally published | Yes |
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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 journal › Article
}
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
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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 -