5-bromo-4-chloro-3-indolyl phosphate (BCIP) is a histochemical substrate for phosphatase. BCIP is used in conjunction with Nitro Blue Tetrazolium (NBT, cat. no. 30550) for the colorimetric detection of alkaline phosphatase-labelled molecules in Northern, Southern, and Western blotting, in situ hybridization, and immunohistochemistry. When incubated with alkaline phosphatase the BCIP/NBT substrate produces an insoluble, purple coloured NBT diformazan precipitate that is easily detected on membranes or tissue sections.BCIP p-toluidine salt and NBT should be solved in dimethylformamide for preparation of a stock solution.Stock solution: 50 mg/ml in 100 % DMF. Store at 4 °C or -20 °C.Staining 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).For added convenience ready-to-use BCIP/NBT solutions are provided: SERVAColor BCIP/NBT Blot Solution, cat. no. 15245 and BCIP/NBT Ready-To-Use Substrate for Immunohistochemistry and Blotting, cat. no. 15246. 5-bromo-4-chloro-3-indolyl phosphate (BCIP) is a histochemical substrate for phosphatase. BCIP is used in conjunction with Nitro Blue Tetrazolium (NBT, cat. no. 30550) for the colorimetric detection of alkaline phosphatase-labelled molecules in Northern, Southern, and Western blotting, in situ hybridization, and immunohistochemistry. When incubated with alkaline phosphatase the BCIP/NBT substrate produces an insoluble, purple coloured NBT diformazan precipitate that is easily detected on membranes or tissue sections.BCIP p-toluidine salt and NBT should be solved in dimethylformamide for preparation of a stock solution.Stock solution: 50 mg/ml in 100 % DMF. Store at 4 °C or -20 °C.Staining 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).For added convenience ready-to-use BCIP/NBT solutions are provided: SERVAColor BCIP/NBT Blot Solution, cat. no. 15245 and BCIP/NBT Ready-To-Use Substrate for Immunohistochemistry and Blotting, cat. no. 15246.