BCIP, molecular biology grade, 250 mg

Substrates

5-Bromo-4-chloro-3-indolyl phosphate p-toluidine salt

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: 1. Add the following components to 10mL of the Alkaline Phosphatase (AP) buffer (100mM Tris-HCl (pH9.5), 100mM NaCl and 10mM MgCl2): 33µL BCIP (50mg/mL in DMF) plus 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.