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B6149 Sigma

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

≥98% (HPLC)

Synonym: BCIP®, X-phosphate disodium salt




1, 5 g in poly bottle

500 mg in poly tube

25, 50, 100 mg in poly bottle


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.

Legal Information

BCIP is a registered trademark of Sigma-Aldrich Co. LLC

Price and Availability

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NBT-BCIP<SUP>®</SUP> solution

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Nitrotetrazolium Blue chloride

powder, electrophoresis grade

Safety & Documentation

Safety Information

WGK Germany 
Protocols & Articles


Colorimetric Alkaline Phosphatase and Peroxidase Substrate Detection Systems

Nitroblue Tetrazolium (NBT) is used with the alkaline phosphatase substrate 5-Bromo- 4-Chloro-3-Indolyl Phosphate (BCIP) in western blotting and immunohistological staining procedures. These substrat...
BioFiles 2008, 3.4, 6.
Keywords: Applications, Enzyme-linked immunosorbent assay, Formulations, Immunohistochemistry, Western blot

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