NBT

Description

• 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.[1]
• Application: Immunohistocytochemistry[2]Western blot[3]Southern blotNorthern blotColony and plaque hybridizationIn situ hybridization[4]
• 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 solutionAlways 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 NaClNote: 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.