BCIP, molecular biology grade

BCIP (5-bromo-4-chloro-3-indolyl phosphate, p-toluidine salt) is a chromogenic substrate for alkaline phosphatase, used in combination with the oxidant NBT (nitro blue tetrazolium) to enhance blue color development.

Applications:
When alkaline phosphatase conjµgates are used for detection, BCIP is recommended for the following applications: Southern blotting, Western blotting, Northern blotting and dot/slot blots.

It is recommended to prepare a 50mg/mL stock solution in DMF.
Protocol for Preparation of BCIP/NBT Substrate Solution:
Add the following components
- 10mL of the Alkaline Phosphatase (AP) buffer (100mM Tris-HCl (pH9.5), 100mM NaCl and 10mM MgCl2)
- 33µL BCIP (50mg/mL in DMF)
- 44µL NBT (75mg/mL in 70% DMF).

NOTE: Avoid exposure to light - Prepare fresh developing solution and use within an hour - BCIP and NBT solutions in DMF do not freeze. They are stable for approximately 2 years when stored at -20°C in the dark.