Product Description

5-bromo, 4-chloro, 3-indolylphosphate (BCIP)/Nitro-Blue Tetrazolium (NBT) substrate, used for the localization of alkaline phosphatase (AP) labeled probes on Western, Northern, Southern and dot blots. BCIP acts as the substrate for AP and reacts further after the dephosphorylation to give a dark-blue indigo-dye as an oxidation product; NBT serves herein as the oxidant and also gives a dark-blue dye, intensifying the color and making the detection more sensitive.

Offered as 10 ml:

Molecular Weight: NBT: MW = 817.7, BCIP toluidine salt: MW = 433.6
Molecular Formula: NBT: ·C40H30Cl2N10O6, ·BCIP: ·C8H6NO4BrClP ·x ·C7H9N
Form: Liquid
Physical Appearance: Yellow to light brown liquid
Concentration: 50X

Preparation of staining solution:
•Add 200 µl of BCIP/NBT stock solution to the following room temperature buffer: 10 ml 0.1 M Tris Hydrochloride, pH 9.5, 0.1 M Sodium Chloride, and 0.05 M Magnesium Chloride. (do not include Magnesium Chloride in the detection buffer, this may cause spotty background on the membrane after the detection procedure)
•Prepare solution immediately prior to use.
•Discard if solution is turbid or purple.
•For Research Use Only

Technical Specifications