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INT/BCIP Stock Solution

solution, pkg of 3 mL

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5-bromo-4-chloro-3-indoyl phosphate, BCIP



Quality Level

mol wt

(INT: Mr = 505.7; BCIP toluidine salt: Mr = 433.6)


pkg of 3 mL



storage temp.


General description

BCIP (5-bromo-4-chloro-3-indoyl phosphate) is the AP (alkaline phosphatase)-substrate which after dephosphorylation reacts further to give a dark-blue indigo dye as an oxidation product. INT (iodonitrotetrazolium) is the oxidant producing a red dye. The reaction product has a reddish-brown color and is insoluble in water.


INT/BCIP Stock Solution is used for the sensitive detection of alkaline phosphatase (AP) in blotting protocols, that includes:
  • Southern blot
  • Western blot
  • immunohistochemistry and immunocytochemistry
It has been used for colorogenic in situ hybridization.


Formulas: INT: C19H13CIN5O2; BCIP: C8H6NO4BrCIP x C7H9N


BCIP is the AP-substrate which after dephosphorylation reacts further to give a dark-blue indigo dye as an oxidation product. INT is the oxidant producing a red dye.

Physical form

Solution of 33 mg/ml INT (2-[4-iodophenyl]-3-[4-nitrophenyl]-5-phenyltetrazolium chloride) and 33 mg/ml BCIP (5-bromo-4-chloro-3-indolyl-phosphate, toluidine-salt in DMSO)

Preparation Note

Working solution: Preparation of 10 ml Staining Solution
Bring the stock solution to 15 to 25 °C until all components are dissolved.
Add 75 μl of the stock solution to 10 ml 0.1 M Tris-buffer, pH 9.5, 0.05 M MgCl2, 0.1 M NaCl.
Note: Prepare the staining solution shortly before use.
Preparation of Additional Solutions Required
Blocking solution: Dissolve 0.5 g Blocking Reagent in 100 ml TBS, pH 7.5, by heating to 50 to 60 °C (1 hour). Dissolving the components can be accelerated by sonication or by incubation in a microwave oven.
Information Note: The solution remains turbid.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Storage Class Code

12 - Non Combustible Liquids



Flash Point(F)

does not flash

Flash Point(C)

does not flash

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