A simple method for the solubilisation of reduced NBT, and its use as a colorimetric assay for activation of human macrophages by gamma-interferon.
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.
D000818:Animals; D003124:Colorimetry / Q000379:methods; D004121:Dimethyl Sulfoxide; D006801:Humans; D006878:Hydroxides; D007371:Interferon-gamma / Q000276:immunology; D008262:Macrophage Activation; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D009000:Monocytes / Q000276:immunology; D009580:Nitroblue Tetrazolium; D010084:Oxidation-Reduction; D011188:Potassium; D017680:Potassium Compounds; D012995:Solubility; D013778:Tetrazolium Salts
D006878:Hydroxides; D017680:Potassium Compounds; D013778:Tetrazolium Salts; D009580:Nitroblue Tetrazolium; D007371:Interferon-gamma; D011188:Potassium; C029943:potassium hydroxide; D004121:Dimethyl Sulfoxide