BCIP/NBT Substrate

Product Description:

The BCIP/NBT mono-component solution is a ready-to-use substrate system containing 5-bromo-4-chloro-3-indolyl-phosphate and nitroblue tetrazolium. This is utilised in the measurement of alkaline phosphatase (AP) probe activity, especially in blotting procedures. The unreacted substrate should be clear and light yellow in appearance. When reacted with alkaline phosphatase, it yields an intense, insoluble black-purple precipitate.

Hazard Identification:

The BCIP/NBT substrate does not contain organic solvents. According to the European Communities legislation on dangerous substances, this product is non-hazardous and does not contain hazardous ingredients at the concentrations used. Refer to the MSDS for additional information.

Storage and Stability:

The BCIP/NBT substrate solution is stable for at least 3 years from date of manufacture if stored at 2-8°C away from light.


  1. Remove the solution from storage and allow to reach room temperature before use.
  2. Wash the specimens incubated with an alkaline phosphatase conjugate in a wash buffer. Do not use phosphate buffers as inorganic phosphate is an inhibitor of AP.
  3. Cover the entire specimen with BCIP/NBT solution for approximately 10 minutes to allow the colour reaction to develop. The length of time required may vary depending on the specimen and the desired optimal signal intensity.
  4. Stop the enzyme reaction by thoroughly washing the specimen with distilled water.


We offer a range of substrates with varying performances. We encourage you to contact us with your requirements so that we can recommend the most suitable product for you.
All substrates can be adapted for use with immunohistochemistry (IHC) upon request.

BCIP/NBT Substrate I (BN-001)

Provides the fastest reaction dynamics, suitable for blotting procedures.

BCIP/NBT Substrate II (BN-002)

Slower reaction dynamics than BN-001, suitable for blotting procedures.

BCIP/NBT Substrate III (BN-003)

Slower reaction dynamics than BN-002, suitable for blotting procedures

BCIP/NBT Substrate IV (BN-004)

Slowest reaction dynamics, suitable for blotting procedures.

Additional documentation:

pdf icon SDS
This SDS is applicable to all Biopanda BCIP/NBT substrates.

This product is for research use only and not intended for use in humans, therapeutic or diagnostic purposes.