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Thermo Scientific™ 1-Step™ NBT/BCIP Substrate Solution

Catalog No. PI34042
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250 mL
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PI34042 250 mL
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Catalog No. PI34042 Supplier Thermo Scientific™ Supplier No. 34042
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Description

Powders and ready-made nitro-blue tetrazolium and 5-bromo-4-chloro-3'-indolyphosphate solutions for chromogenic detection of alkaline phosphatase probes.

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

1-Step™ NBT/BCIP

  • Ideal for immunoblotting applications
  • Minimal assay-to-assay variability

1-Step NBT/BCIP Plus Suppressor

  • Ideal for staining tissues assayed with calf intestinal alkaline phosphatase without the interference caused by endogenous phosphatases
  • Contains 1mM levamisole to inhibit endogenous phosphatase activity

Specifications

Description Pierce 1-Step NBT/BCIP Substrate Solution
Product Type Substrate
Detection Method Colorimetric
Form Liquid
Membrane Compatibility Nitrocellulose, PVDF
Product Line 1-Step
Quantity 250 mL
Substrate Alkaline Phosphatase
Reactivity Alkaline Phosphatase
Content And Storage Upon receipt store product at 4°C. Product is shipped at ambient temperature.

Frequently Asked Questions (FAQs)

What alkaline phosphatase substrates do you offer?

For colorimetric alkaline phosphatase substrates, we offer the following products:

Novex AP Chromogenic Substrate (BCIP/ NBT) (Cat. No. WP20001)

1-Step NBT/BCIP Substrate Solution (Cat. No. 34042)

1-Step PNPP Substrates Solution (Cat. No. 37621)

NBT and BCIP are also available as standalone products:

NBT Substrate Powder (nitro-blue tetrazolium chloride) (Cat. No. 34035)

BCIP Substrate Powder (5-bromo-4-chloro-3'-indolylphosphate p-toluidine) (Cat. No. 34040)

For chemiluminescent alkaline substrates, we offer the following products:

Novex AP Chemiluminescent Substrate (CDP-Star) (Cat. No. WP20002)

NovaBright Phospha-Light EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection (Cat. Nos. N10577, N10578)
Will the 1-Step NBT/BCIP Substrate Solution work with PVDF membranes?

Yes, it will work with both PVDF as well as nitrocellulose membranes
With the 1-Step NBT/BCIP Substrate Solution, how can I eliminate endogenous phosphatase activity?

To eliminate endogenous phosphatase activity, add a final concentration of 1 mM levamisole to the 1-Step NBT/BCIP Substrate Solution.
Can I use the 1-Step NBT/BCIP Substrate Solution for ELISA?

No, the 1-Step NBT/BCIP Substrate Solution cannot be used for ELISA. It is a precipitating alkaline phoshpatase substrate developed for immunoblotting, immunohistochemistry and in situ hybridzation. For ELISA, use our 1-Step PNPP Substrate Solution (Cat. No. 37621).

For Research Use Only. Not for use in diagnostic procedures.