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5-Bromo-4-chloro-3-indolyl phosphate p-toluidine salt


BCIP p-toluidine salt, X-phosphate p-toluidine salt
Empirical Formula (Hill Notation):
C8H6BrClNO4P · C7H9N
CAS Number:
Molecular Weight:
EC Number:
MDL number:
PubChem Substance ID:



Quality Level

storage temp.


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InChI key


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Substrate of choice for use with alkaline phosphatase in immunoblotting and, less commonly, in immunohistological staining procedures. High assay sensitivity is achieved via amplification when BCIP is used in conjunction with Nitro Blue Tetrazolium (NBT) Tablets (Catalog No. N5514). This substrate produces an insoluble end product that is blue-purple in color and can be observed visually.


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.


10, 25 tablets in glass bottle


Contains 25 mg substrate per tablet.


Histochemical substrate for alkaline phosphatase.

Legal Information

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

Storage Class Code

13 - Non Combustible Solids



Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificate of Analysis

Certificate of Origin

More documents

Quotes and Ordering

Substrates for cytochemical demonstration of enzyme activity. II. Some dihalo-3-indolyl phosphates and sulfates.
J P Horwitz et al.
Journal of medicinal chemistry, 9(3), 447-447 (1966-05-01)
M S Blake et al.
Analytical biochemistry, 136(1), 175-179 (1984-01-01)
A rapid, sensitive method has been developed to detect antibody-antigen complexes on "Western blots." The methods of H. Towbin, T. Staehlin, and J. Gordon were used to separate and blot the antigens onto nitrocellulose. The remaining sites of attachment were
Karuppiah Muthumani et al.
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Fernando Cartón-García et al.
Scientific reports, 5, 12312-12312 (2015-07-24)
Inherited MYO5B mutations have recently been associated with microvillus inclusion disease (MVID), an autosomal recessive syndrome characterized by intractable, life-threatening, watery diarrhea appearing shortly after birth. Characterization of the molecular mechanisms underlying this disease and development of novel therapeutic approaches
Dorota Rybaczek et al.
Histochemistry and cell biology, 135(3), 263-280 (2011-02-25)
Here, we demonstrate that in HeLa cells, Ser317 of Chk1 undergoes phosphorylation in response to replication stress induced by hydroxyurea. We also demonstrate the existence of constitutive (interphase and mitotic) Chk1 kinase phosphorylation, the translocation of its phosphorylated form from


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 substrate systems produce an insoluble NBT diformazan end product that is blue to purple in color and can be observed visually.

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