5-bromo-4-chloro-3-indolyl phosphate (BCIP) is a histochemical substrate for phosphatase. BCIP is used in conjunction with Nitro Blue Tetrazolium (NBT, cat. no. 30550) for the colorimetric detection of alkaline phosphatase-labelled molecules in Northern, Southern, and Western blotting, in situ hybridization, and immunohistochemistry. When incubated with alkaline phosphatase the BCIP/NBT substrate produces an insoluble, purple coloured NBT diformazan precipitate that is easily detected on membranes or tissue sections.
BCIP p-toluidine salt and NBT should be solved in dimethylformamide for preparation of a stock solution.
Stock solution: 50 mg/ml in 100 % DMF. Store at 4 °C or -20 °C.
Staining solution for Western blots: 66 µl NBT stock solution and 33 µl BCIP stock solution in 10 ml staining buffer (100 mM NaCl, 5 mM MgCl₂, 100 mM Tris; pH 9.5).
For added convenience ready-to-use BCIP/NBT solutions are provided: SERVAColor BCIP/NBT Blot Solution, cat. no. 15245 and BCIP/NBT Ready-To-Use Substrate for Immunohistochemistry and Blotting, cat. no. 15246.

Assay (HPLC)

min. 99.0 %

References:

1. Wolf, P.L. et al. (1973) Clin. Chem. 19, 1248-9

EINECS: 229-506-1 • WGK: 1 • HS: 29339980
Storage Temperature: -15 °C to -25 °C