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5-Bromo-4-chloro-3-indolyl phosphate

BCIP
IUPAC name 5-Bromo-4-chloro-3-indolyl phosphate
Other names BCIP
Properties
Molecular formula C8H6BrClNO4P
Molar mass 433.6 g/mol
Appearance Colorless
Solubility in water very low solubility in water or lipid
Except where noted otherwise, data are given for
materials in their standard state
(at 25 °C, 100 kPa)
Infobox disclaimer and references

5-Bromo-4-chloro-3-indolyl phosphate (BCIP) is a chemical compound used in immunoblotting, with nitro blue tetrazolium chloride (NBT), for sensitive detection of alkaline phosphatase. NBT serves as the oxidant (and gives also dark blue dye) and BCIP is the alkaline phosphatase substrate. Alkaline phosphatase is commonly conjugated to secondary antibodies.

BCIP (colorless)oxidation→ blue precipitate.

BCIP-NBT naturally forms this bluish purple precipitate over time; however, alkaline phosphatase speeds up the process 1000 fold. BCIP binds very tightly in the alkaline phosphatase active site, but when NBT reacts with BCIP, it is released from the enzyme and the colored precipitate forms.

References


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