All Photos(1)



BCIP®/NBT-Purple Liquid Substrate System for Membranes

ready to use alkaline phosphatase substrate


Quality Level



storage temp.



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


Exclamation mark

Signal Word


Hazard Statements

Hazard Classifications

Skin Sens. 1

Storage Class Code

10 - Combustible liquids



Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

Xudong Wang et al.
Frontiers in cell and developmental biology, 9, 690502-690502 (2021-08-28)
Melatonin (Mlt) confers potential antitumor effects in various types of cancer. However, to the best of our knowledge, the role of Mlt in the giant cell tumor of bone (GCTB) remains unknown. Moreover, further research is required to assess whether
pSTAT5 and ERK exhibit different expression in myeloproliferative neoplasms
Wisniewska-Chudy E, et al.
Oncology Reports, 37(4), 2295-2307 (2017)
Characterization of the 41 kDa allergen Asp v 13, a subtilisin-like serine protease from Aspergillus versicolor
Shi C and Miller JD
Molecular Immunology, 48(15-16), 1827-1834 (2011)
Membrane bioreactor separator system for integrated IgG fragmentation and Fab purification
Yu D and Ghosh R
Journal of Immunological Methods, 359(1-2), 37-41 (2010)
Louise Bjerkan et al.
Frontiers in immunology, 12, 720550-720550 (2021-11-05)
Targeted delivery of antigen to antigen presenting cells (APCs) is an efficient way to induce robust antigen-specific immune responses. Here, we present a novel DNA vaccine that targets the Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5), a leading blood-stage antigen


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.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service