The ditetrazolium compound nitro blue tetrazolium chloride, also known as NBT for short, serves as a substrate for the detection of succinate dehydrogenase (1), glucose-6-phosphate dehydrogenase (2) and xanthine oxidase (3). NBT is used in conjunction with BCIP (cat. no. 15247) for the colourimetric detection of alkaline phosphatase labelled molecules in Northern, Southern and Western blotting, in situ hybridisation and immunohistochemistry. When incubated with alkaline phosphatase, the BCIP/NBT substrate produces an insoluble, violet-stained NBT-diformazan precipitate that is easily detected on membranes or tissue sections.BCIP p-toluidine salt and NBT should be dissolved in dimethylformamide to prepare a stock solution.For added convenience, ready-to-use BCIP/NBT solutions are offered: SERVAColor BCIP/NBT Blot Solution, cat. no. 15245 and BCIP/NBT Ready-to-use Substrate for Immunohistochemistry and Blotting, cat. no. 15246.Stock solution for Western blots: 66 µl NBT stock solution and 33 µl BCIP stock solution in 10 ml staining buffer (100 mM NaCl, 5 mM MgCl2, 100 mM Tris, pH 9.5). The ditetrazolium compound nitro blue tetrazolium chloride, also known as NBT for short, serves as a substrate for the detection of succinate dehydrogenase (1), glucose-6-phosphate dehydrogenase (2) and xanthine oxidase (3). NBT is used in conjunction with BCIP (cat. no. 15247) for the colourimetric detection of alkaline phosphatase labelled molecules in Northern, Southern and Western blotting, in situ hybridisation and immunohistochemistry. When incubated with alkaline phosphatase, the BCIP/NBT substrate produces an insoluble, violet-stained NBT-diformazan precipitate that is easily detected on membranes or tissue sections.BCIP p-toluidine salt and NBT should be dissolved in dimethylformamide to prepare a stock solution.For added convenience, ready-to-use BCIP/NBT solutions are offered: SERVAColor BCIP/NBT Blot Solution, cat. no. 15245 and BCIP/NBT Ready-to-use Substrate for Immunohistochemistry and Blotting, cat. no. 15246.Stock solution for Western blots: 66 µl NBT stock solution and 33 µl BCIP stock solution in 10 ml staining buffer (100 mM NaCl, 5 mM MgCl2, 100 mM Tris, pH 9.5).