Application

For the colorimetric detection of alkaline phosphatase-labeled molecules, 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) and Nitro Blue Tetrazolium (NBT) are available. The BCIP/NBT substrate system is versatile and functions in a variety of applications, including Northern Southern, and Western blotting, in situ hybridization, and immunohistochemistry. BCIP is provided in two salt forms: the disodium salt which is soluble in water and the p-toluidine form which is soluble in dimethylformamide. These salt forms may be used to prepare a stock solution that in combination with NBT and a reaction buffer, form a substrate solution for alkaline phosphatase. This substrate system, when incubated with alkaline phosphatase, produces an insoluble NBT diformazan product that is easily observable with its purple color. For added convenience, a mixture of BCIP and NBT is provided in an easily dissolvable tablet form or as a ready-to-use liquid.

Legal Information

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

Packaging

10, 25 tablets in glass bottle

Quantity

Contains 25 mg substrate per tablet.

Specificity

Substrate of choice for use with alkaline phosphatase in immunoblotting and, less commonly, in immunohistological staining procedures. High assay sensitivity is achieved via amplification when BCIP is used in conjunction with Nitro Blue Tetrazolium (NBT) Tablets (Catalog No. N5514). This substrate produces an insoluble end product that is blue-purple in color and can be observed visually.

Substrates

Histochemical substrate for alkaline phosphatase.