Thermo Scientific™

NBT Substrate Powder (nitro-blue tetrazolium chloride)

Catalog number:  34035

Related applications:

Cellular Imaging

| Protein Electrophoresis & Western Blotting

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Thermo Scientific Pierce NBT and BCIP Substrates are powders and ready-made solutions of nitro-blue tetrazolium and 5-bromo-4-chloro-3'-indolyphosphate for chromogenic blot and IHC staining with alkaline phosphatase probes.

The combination of NBT (nitro-blue tetrazolium chloride) and BCIP (5-bromo-4-chloro-3'-indolyphosphate p-toluidine salt) yields an intense, insoluble black-purple precipitate when reacted with alkaline phosphatase, a popular enzyme conjugate for antibody probes. Pierce NBT and BCIP powders are available individually and in two convenient, ready-to-use solutions (with and without a levamisole suppressor of endogenous phosphatase activity).


AP substrate—for detection of alkaline phosphatase (alk-phos) activity on solid media, including nitrocellulose and PVDF membranes and fixed tissue samples
Chromogenic—no special equipment needed for visualization; produces purple-black precipitate that is easy to photograph
Package options—choose individual NBT and BCIP powders, or preformulated, ready-to-use solutions with or without levamisole suppressor

Highlights of Pierce NBT/BCIP 1-Step Solution (Part No. 34042):
• Best for immunoblotting applications
• Ready-to-use single component
• Sensitive with low background and minimial assay-to-assay variability

Highlights of Pierce NBT/BCIP plus Suppressor 1-Step Solution (Part No. 34070):
• Best for immunohistochemistry applications
• Ready-to-use single component
• Contains 1 mM levamisole to inhibit endogenous phosphatase activity, while yielding an intense dark purple precipitate in the presence of calf intestinal phosphatase

Related Products
1-Step™ NBT/BCIP Substrate Solution
1-Step™ NBT/BCIP plus Suppressor Substrate Solution
BCIP Substrate Powder (5-bromo-4-chloro-3'-indolyphosphate p-toluidine)
For Research Use Only. Not for use in diagnostic procedures.


Detection Method: Colorimetric
Substrate Properties: Chemical Substrate
Substrate Type: AP (Alkaline Phosphatase) Substrate
Description: Pierce NBT Substrate

Contents & storage

Upon receipt store product at 4°C protected from moisture. Product is shipped at ambient temperature.