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  • B3679 - BCIP®/NBT-Purple Liquid Substrate System for Membranes

B3679 Sigma-Aldrich

BCIP®/NBT-Purple Liquid Substrate System for Membranes

ready to use alkaline phosphatase substrate

  •  NACRES NA.83




BCIP®/NBT-Purple Liquid Substrate System for Membranes has been used as a substrate for alkaline phosphatase
• for the detection of secondary antibodies by immunoblot
• in ELISPOT assay
• in dot blot analysis
• in western blot to visualize protein bands on nitrocellulose membrane

Biochem/physiol Actions

5-bromo-4-chloro-3-indolyl phosphate (BCIP)/ nitro blue tetrazolium (NBT) is a chromogenic substrate system for the detection of alkaline phosphatase (AP) enzyme. AP hydrolyses BCIP to form an intermediate leucoindigo compound and is further oxidized by NBT to form 5,5′-dibromo-4,4′-dichloro indigo (BCI). NBT is reduced to an insoluble purple diformazan (DF) and forms NBT-DF. A mixture of NBT-DF and BCI results in the dark purple colored precipitate. AP by hydrolysing many BCIP molecules makes it a highly sensitive method.


Stable at least one year at 2-8 °C.

Physical form


Legal Information

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

Safety & Documentation

Safety Information

GHS07  GHS07
Signal word 
Hazard statements 
NONH for all modes of transport
WGK Germany 


Certificate of Analysis (COA)

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Certificate of Origin (COO)

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Single- & multi-channel pipettes from BRAND
Protocols & Articles


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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: Enzyme-linked immunosorbent assay, Immunohistochemistry, Western blot

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