Application

BCIP^®/NBT-Purple Liquid Substrate System for Membranes has been used as a substrate for alkaline phosphatasefor the detection of secondary antibodies by immunoblotin ELISPOT assayin dot blot analysisin western blot to visualize protein bands on nitrocellulose membrane

Biochem/physiol Actions

5-bromo-4-chloro-3-indolyl phosphate (BCIP)/ nitro blue tetrazolium (NBT) is a chromogenic substrate system for the detection of alkaline phosphatase (AP) enzyme. AP hydrolyses BCIP to form an intermediate leucoindigo compound and is further oxidized by NBT to form 5,5′-dibromo-4,4′-dichloro indigo (BCI). NBT is reduced to an insoluble purple diformazan (DF) and forms NBT-DF. A mixture of NBT-DF and BCI results in the dark purple colored precipitate. AP by hydrolysing many BCIP molecules makes it a highly sensitive method.

Caution

Stable at least one year at 2-8 °C.

Legal Information

BCIP is a registered trademark of Sigma-Aldrich Co. LLC

Physical form

Ready-to-use.