NBT SOLUTION

General description

NBT is a chromogenic and soluble substance. It is a substrate of alkaline phosphatase (AP) and is converted to an indigo/purple dye on its action.

Application

• Immunohistocytochemistry
• Western blot
• Southern blot
• Northern blot
• Colony and plaque hybridization
• In situ hybridization

Physical form

Solution (100 mg/ml) in 70% dimethylformamide (DMF) (v/v). The color of the NBT solution can vary between yellow and brown depending on the lot used.

Note: The color does not impair the quality or the function of the reagent. In some cases a precipitate may occur which is easily brought into solution by briefly warming the substrate at 37 °C.

Preparation Note

Working solution: Preparation of staining solution

• Always prepare this solution fresh shortly before use!For all applications except DIG System: Add 50 μl NBT solution and 37,5 μl BCIP 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 50 μl NBT solution and 37,5 μl BCIP to 10 ml 0.1 M Tris-HCl, pH 9.5 (20 °C), 0.1 M NaCl
• Note: Do not include MgCl₂ in the DIG detection buffer as this might lead to spotty background on the membrane after the detection procedure. Alkaline Phosphatase does not require Mg²⁺.

Other Notes

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