AN IMPROVED COLORIMETRIC ASSAY FOR INTERLEUKIN-2
AN IMPROVED COLORIMETRIC ASSAY FOR INTERLEUKIN-2
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DOI：
10.1016/0022-1759(86)90183-3
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发表时间：
1986-01-01
影响因子：
2.2
通讯作者：
TSUKAMOTO, K
中科院分区：
文献类型：
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作者：
TADA, H;SHIHO, O;TSUKAMOTO, K
Mosmann用四氮唑盐3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑（MTT）测定活细胞数的方法经过改进，使其能够一次测定大量的白细胞介素2 （IL-2）样品，减少劳动，提高准确性。对该方法的每一步都进行了详细的检查和修改（修改后的MTT法）。(1)以il -2依赖性小鼠自然杀伤细胞系NKC3作为指示细胞系。(2)用10%十二烷基硫酸钠-0.01盐酸溶液溶解所制得的MTT甲醛酰胺。我们将改进的MTT法与传统的[3H]胸苷嘧啶法（3H- tdr法）得到的值进行了比较，证实IL-2含量的估计值几乎相等。两种方法测定的IL-2含量的标准误差在5%以内。
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). (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 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.