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Sigma-Aldrich

BCIP®/NBT

alkaline phosphatase substrate, chromogenic, liquid

Synonym(s):

5-bromo-4-chloro-3-indolyl-phosphate / Nitro blue tetrazolium chloride solution, BCIP/NBT Alkaline Phosphatase Substrate solution

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About This Item

UNSPSC Code:
12352204
NACRES:
NA.83

product name

BCIP®/NBT Liquid Substrate System, ready to use solution

Quality Level

300

form

liquid

storage temp.

2-8°C

Application

A ready-to-use, precipitating substrate system for alkaline phosphatase. This substrate system produces an insoluble NBT diformazan end product that is blue to purple in color and can be observed visually. Immunohistochemical staining with BCIP/NBT utilizes requires organic mounting media and can be counterstained with Nuclear Fast Red or Light Green. BCIP/NBT is generally a more sensitive method than Fast Red. Not recommended for ELISA (microwell) procedures.
BCIP®/NBT Liquid Substrate System has been used:
  • for staining alkaline phosphatase(ALP) in bone marrow stromal cells
  • as a substrate in western blot detection of matrix metalloproteinase-2 specific antibody
  • as a substrate for streptavidin-alkaline phosphatase in the enzyme-linked immunospot (ELISpot) assays

Caution

It is recommended to store the product between 4 - 8 °C. Do not freeze. The product may be stored at room temperature for up to one year. If a precipitate forms, the solution can be gently swirled or filtered.

Legal Information

BCIP is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Commensal microbes induce serum IgA responses that protect against polymicrobial sepsis
Wilmore JR, et al.
Cell host & microbe, 23(3), 302-311 (2018)
Nana H Overgaard et al.
Vaccine, 35(42), 5629-5636 (2017-09-10)
The relationship between the antigen dose and the quality of an immune response generated upon immunization is poorly understood. However, findings show that the immune system is indeed influenced by the antigen dose; hence underlining the importance of correctly determining
Epigenetic regulation of Runx2 transcription and osteoblast differentiation by nicotinamide phosphoribosyltransferase
Ling M, et al.
Cell & Bioscience, 7(1), 27-27 (2017)
Salma E Jiménez-Badillo et al.
Scientific reports, 7(1), 13351-13351 (2017-10-19)
The Catsper1 gene, whose expression is restricted to male germ cells, has great importance in reproductive biology because of its function in sperm motility and fertilization. We previously reported that the promoter of this gene has transcriptional activity in either
Yi Sun et al.
Veterinary research, 42, 62-62 (2011-05-17)
Babesia divergens is a tick-transmitted apicomplexan parasite for which asexual multiplication in its vertebrate hosts is restricted to erythrocytes. Current knowledge of invasion of these target cells is limited. An efficient in vitro invasion assay was set up to gain
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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 substrate systems produce an insoluble NBT diformazan end product that is blue to purple in color and can be observed visually.

Protocols

Immunodetection Using BCIP/NBT Substrate

5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium (NBT) is an ideal insoluble substrate for use with alkaline phosphatase. This system produces a blue-purple product. The intense color can be observed visually, is very stable, and will not fade upon exposure to light.

10x Blocking Buffer Protocol

Membrane-based blotting applications that employ enzyme conjugates to generate colorimetric or chemiluminescent signal require the use of an added blocking step to decrease the signal generated by non-specific binding.

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