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一种溶解还原型氮蓝四唑(NBT)的简单方法及其作为γ干扰素激活人巨噬细胞的比色测定法的应用。

A simple method for the solubilisation of reduced NBT, and its use as a colorimetric assay for activation of human macrophages by gamma-interferon.

作者信息

Rook G A, Steele J, Umar S, Dockrell H M

出版信息

J Immunol Methods. 1985 Sep 3;82(1):161-7. doi: 10.1016/0022-1759(85)90235-2.

Abstract

We describe a simple method by which the insoluble blue formazan dye produced by the reduction of nitro blue tetrazolium can be dissolved without heating using potassium hydroxide and dimethyl sulphoxide. This modification enhances the sensitivity and increases the applications of tests performed using the microELISA method and removes variations caused by uneven cell monolayers. It also allows quantification of NBT reduced by cells adherent to coverslips or in larger wells or Petri dishes, and can be used as a sensitive assay for macrophage activation by gamma-interferon.

摘要

我们描述了一种简单的方法,通过该方法,还原硝基蓝四氮唑产生的不溶性蓝色甲臜染料可以在不加热的情况下用氢氧化钾和二甲基亚砜溶解。这种改进提高了灵敏度,增加了使用微量酶联免疫吸附测定法进行的测试的应用范围,并消除了由不均匀细胞单层引起的变化。它还允许对附着在盖玻片上或较大孔或培养皿中的细胞还原的NBT进行定量,并且可以用作γ-干扰素激活巨噬细胞的灵敏测定法。

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