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NBT/BCIP Stock Solution

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



Quality Level

mol wt



pkg of 8 mL



storage temp.


General description

The reaction product has a blue color and is insoluble in water. The blue color can be visually seen.

Formulas: NBT: C40H30Cl2N10O6, BCIP: C8H6NO4BrClP x C7H9N

Evaluation: visually


  • The solution is used for the sensitive detection of alkaline phosphatase (AP) in blotting protocols, including: Southern blot
  • Northern blot
  • Western blot[1]
  • Colony and plaque lifts
  • In situ hybridization[2][3]
  • and for applications in immunohistochemistry and immunocytochemistry.
Note: NBT/BCIP may be used with both nitrocellulose and nylon membranes.

Physical form

Solution of 18.75 mg/ml nitro blue tetrazolium chloride and 9.4 mg/ml 5-bromo-4-chloro-3-indolyl-phosphate, toluidine-salt in 67% (DMSO) (v/v).
Note: The color of the solution can vary from yellowish to light brown.

Preparation Note

Working solution: Preparation of staining solution
Always prepare this solution fresh shortly before use.
  • For all applications except DIG System: Add 200 μl of the stock solution to 10 ml 0.1 M Tris-HCl, pH 9.5 (20 °C), 0.1 M NaCl, 0.05 M MgCl2.
  • For DIG System applications: Add 200 μl of the stock solution to 10 ml 0.1 M Tris-HCl, pH 9.5 (20 °C), 0.1 M NaCl
Note: Do not include MgCl2 in the detection buffer as this may lead to spotty background on the membrane after the detection procedure. Insoluble in water.

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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