5-Bromo-4-chloro-3-indolyl phosphate disodium salt 

Histochemical substrate for alkaline phosphatase.

For the colorimetric detection of alkaline phosphatase-labeled molecules, 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) and Nitro Blue Tetrazolium (NBT) are available. The BCIP/NBT substrate system is versatile and functions in a variety of applications, including Northern Southern, and Western blotting, in situ hybridization, and immunohistochemistry.

BCIP is provided in two salt forms: the disodium salt which is soluble in water and the p-toluidine form which is soluble in dimethylformamide. These salt forms may be used to prepare a stock solution that in combination with NBT and a reaction buffer, form a substrate solution for alkaline phosphatase. This substrate system, when incubated with alkaline phosphatase, produces an insoluble NBT diformazan product that is easily observable with its purple color. For added convenience, a mixture of BCIP and NBT is provided in an easily dissolvable tablet form or as a ready-to-use liquid.

BCIP is a registered trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.

for molecular biology

≥98%

powder

H₂O: 20 mg/mL

DMF: insoluble

Protease, none detected

−20°C

McGadey, J., A tetrazolium method for non-specific alkaline phosphatase. Histochemie 23, 180-184, (1970) abstract

Knecht, D.A., Dimond, R.L., Visualization of antigenic proteins on Western blots. Anal. Biochem. 136, 180, (1984)

Leary, J.J., et al., Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc. Natl. Acad. Sci. USA 80, 4045-4049, (1983)

Horwitz, J.P., et al. J. Med. Chem. 9, 447, (1966)

Walters, C., et al., Detection of parvovirus B19 in macerated fetal tissue using in situ hybridization. J. Clin. Pathol. 50(9), 749-754, (1997)

Meltzer, J.C., et al., Enhanced immunohistochemical detection of autonomic nerve fibers, cytokines and inducible nitric oxide synthase by light and fluorescent microscopy in rat spleen. J. Histochem. Cytochem. 45(4), 599-610, (1997)

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