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
Note: NBT/BCIP may be used with both nitrocellulose and nylon membranes.
- The solution is used for the sensitive detection of alkaline phosphatase (AP) in blotting protocols, including: Southern blot
- Northern blot
- Western blot
- Colony and plaque lifts
- In situ hybridization
- and for applications in immunohistochemistry and immunocytochemistry.
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.
Working solution: Preparation of staining solution
Always prepare this solution fresh shortly before use.
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.
- 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
For life science research only. Not for use in diagnostic procedures.