BCIP (5-bromo-4-chloro-3-indolyl-phosphate) is used in conjunction with NBT (nitro blue tetrazolium) for the colorimetric detection of alkaline phosphatase activity. Each vial of BCIP is supplied with a vial of NBT.
Preparation of Substrates to Detect Alkaline Phosphatase: For every 5ml of alkaline phosphatase buffer (100mM Tris-HCl [pH 9.0], 150mM NaCl, 1mM MgCl2), add 33μl NBT and 16.5μl BCIP. Add the NBT first, mix, add the BCIP, and mix again. Use within 1 hour, and discard any unused solution.
Concentration: BCIP (50mg/ml) in 100% dimethylformamide; NBT (50mg/ml) in 70% dimethylformamide....
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Concentration: BCIP (50mg/ml) in 100% dimethylformamide; NBT (50mg/ml) in 70% dimethylformamide.
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BCIP/NBT Color Development Substrate
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