NBT/BCIP Stock Solution

Overview

Suitable as substrate for alkaline phosphatase detection in applications such as IHC & immunoblotting (e.g. Dot blot). Produces an insoluble, blue- purple end product following reaction with alkaline phosphatase (AP).

Contents

Solution of 18.8 mg/ml nitro-blue tetrazolium chloride; 9.4 mg/ml 5-bromo-4-chloro-3-indolylphosphate toluidine salt in 67% DMSO.

Preparation note - preparation of staining solution

Always prepare fresh solutions, 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 NaCl

Note: Please do not include MgCl2 in the detection buffer as this can lead to spotty background on the membrane after the detection procedure. Insoluble in water.