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11681451001 Roche

NBT/BCIP Stock Solution



Related Categories Colorimetric, Molecular Biology, Nucleic Acid Detection Substrates and Reagents, Nucleic Acid Detection and Hybridization
Quality Level   100
form   solution
mol wt   817.7
packaging   pkg of 8 mL
mfr. no.   Roche
shipped in   wet ice
storage temp.   2-8°C


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 C7H9N

Evaluation: visually


• The solution is used for the sensitive detection of alkaline phosphatase (AP) in blotting protocols, including: Southern blot
• Northern blot
• Western blot
• Colony and plaque lifts
• In situ hybridization
• and for applications in immunohistochemistry and immunocytochemistry.
Note: NBT/BCIP may be used with both nitrocellulose and nylon membranes.

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 solution
Always 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 NaCl
Note: 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.

Other Notes

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

Safety & Documentation

Safety Information

NONH for all modes of transport


Certificate of Analysis (COA)

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Protocols & Articles


BCIP Protocol - Technical Article

BCIP: Suitable solvents and storage The recommended solvent will depend on the type of BCIP (5-bromo-4-chloro-3-indolyl-phosphate) salt you are using: The disodium salt of BCIP (Cat.No. 1 017 373) is...
Keywords: Filtration, Precipitation, Solvents

NBT/BCIP Stock Solution Protocol & Troubleshooting

There are several counterstains possible in combination with BM Purple (or NBT/BCIP in general), for instance, FastGreen FCF or Nuclear Fast Red. For details, see the protocol described in the Roche ...
Keywords: In Situ hybridization, Nucleic acid hybridization

Peer-Reviewed Papers


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