Not available outside of the UK & Ireland.

Application

The solution is used for the sensitive detection of alkaline phosphatase (AP) in blotting protocols, including: Southern blotNorthern blotWestern blotColony and plaque liftsIn situ hybridizationand for applications in immunohistochemistry and immunocytochemistry.Note: NBT/BCIP may be used with both nitrocellulose and nylon membranes.

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 C₇H₉NEvaluation: visually

Other Notes

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

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 solutionAlways 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 NaClNote: 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.