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B3679

Sigma-Aldrich

BCIP®/NBT-Blue

alkaline phosphatase substrate, chromogenic, liquid

Synonym(s):

5-bromo-4-chloro-3-indolyl-phosphate / Nitro blue tetrazolium chloride liquid substrate system

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About This Item

UNSPSC Code:
12352204
NACRES:
NA.83

product name

BCIP®/NBT-Purple Liquid Substrate System for Membranes, ready to use alkaline phosphatase substrate

Quality Level

100

form

liquid

storage temp.

2-8°C

Related Categories

Application

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.

Caution

Stable at least one year at 2-8 °C.

Physical form

Ready-to-use.

Legal Information

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

pictograms

Exclamation mark
GHS07

signalword

Warning

hcodes

H317

Hazard Classifications

Skin Sens. 1

Storage Class

10 - 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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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

RNAi In Situ

In Situ Hybridization of Whole-Mount Mouse Embryos with RNA Probes: Hybridization, Washes, and Histochemistry. This is a protocol describing how to perform in situ hybridization on whole mouse embryos. Here we describe the hybridization procedure, and the localization of the DIG-labeled RNA using a conjugate of anti-DIG Fab antibody and calf intestinal alkaline phosphatase. Enzyme activity of the reporter is detected by a color reaction, resulting in the formation of a water-insoluble purple/blue precipitate. Manipulating the Mouse Embryo - Third Edition

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