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).
Highlights:
- 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 1mM levamisole to inhibit endogenous phosphatase activity, while yielding an intense dark purple precipitate in the presence of calf intestinal phosphatase
Product Details:
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| Chemical reaction of NBT and BCIP substrates with alkaline phosphatase. BCIP is hydrolyzed by alkaline phosphatase to form an intermediate that undergoes dimerization to produce an indigo dye. The NBT is reduced to the NBT-formazan by the two reducing equivalents generated by the dimerization. This reaction proceeds at a steady rate, allowing accurate control of the relative sensitivity and control of the development of the reaction. |
Product Reference:
- Stuyver, L.J., et al. (2003). Antimicrob. Agents Chemother. 47, 244-254.
Related Resources:
Protein Methods Library: Overview of Immunohistochemistry (IHC)
Protein Methods Library: Immunodetection Strategies for IHC
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