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Pierce NBT/BCIP Substrates and Solutions


Produce blue-purple precipitates for Western blotting or staining applications.

Pierce NBT and BCIP Substrates

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:

Chemical reaction of NBT/BCIP substrate with alkaline phosphatase
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:

  1. Stuyver, L.J., et al. (2003). Antimicrob. Agents Chemother. 47, 244-254.

Related Resources:

Immunohistochemistry (IHC) technical guide
IHC immunodetection technical guide


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
34035 NBT Powder
Formulation: Nitro-blue tetrazolium chloride powder
Sufficient For: 3000 uses at 1X (0.033mg/mL) and 10mL each
1g Product Instructions for product #34035 NBT Powder MSDS for product #34035 NBT Powder Certificate of Analysis for product #34035 NBT Powder $132.00

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34040 BCIP Powder
Formulation: 5-Bromo-4-chloro-3'-indolyphosphate p-toluidine salt powder (≥ 99% pure)
Sufficient For: 6000 uses at 1X (0.017mg/mL) and 10mL each
1g Product Instructions for product #34040 BCIP Powder MSDS for product #34040 BCIP Powder Certificate of Analysis for product #34040 BCIP Powder $221.00

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34042 1-Step NBT/BCIP Solution
Formulation: Proprietary solution of NBT and BCIP
Sufficient For: 25 uses at 10mL each
250mL Product Instructions for product #34042 1-Step NBT/BCIP Solution MSDS for product #34042 1-Step NBT/BCIP Solution Certificate of Analysis for product #34042 1-Step NBT/BCIP Solution $128.00

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34042B 1-Step NBT/BCIP Solution
Formulation: Proprietary solution of NBT and BCIP
Custom MSDS for product #34042B 1-Step NBT/BCIP Solution Certificate of Analysis for product #34042B 1-Step NBT/BCIP Solution
34070 1-Step NBT/BCIP plus Suppressor Solution
Formulation: Proprietary solution of NBT, BCIP and levamisole
Sufficient For: 10 uses at 10mL each
100mL Product Instructions for product #34070 1-Step NBT/BCIP plus Suppressor Solution MSDS for product #34070 1-Step NBT/BCIP plus Suppressor Solution Certificate of Analysis for product #34070 1-Step NBT/BCIP plus Suppressor Solution $95.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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