BCIP is the alkaline phosphatase (AP) substrate, which after dephosphorylation is oxidized by NBT to yield a dark-blue indigo precipitating dye. NBT is thereby reduced to a dark-blue precipitating dye and serves to intensify the color reaction making the detection more sensitive.
The reaction product has a blue color and is insoluble in water. The blue color can be visually seen.
NBT/BCIP Ready-to-Use tablets has been used in:
Note: NBT/BCIP may be used with both nitrocellulose and nylon membranes.
- in situ hybridization.
- western blot
- southern blot
- northern blot
- colony and plaque lifts
Working solution: Preparation of Color Substrate Solution
Add 1 tablet to 10 ml double-dist. water to prepare a ready-to-use staining solution.
Composition of staining solution (pH 9.5) after addition of double-dist. water: 0.4 mg/ml NBT; 0.19 mg/ml BCIP; 100 mM Tris buffer, pH 9.5; 50 mM MgSO4
Information Note: The color of the solution can vary from slightly yellowish-violet to light brown.
Important Note: Prepare the solution shortly before use.
For life science research only. Not for use in diagnostic procedures.