Thermo Scientific™ 1-Step™ NBT/BCIP Substrate Solution (nitro-blue tetrazolium chloride)

The Thermo Scientific™ Pierce NBT and BCIP Substrates are designed for chromogenic blotting and immunohistochemistry (IHC) applications. These substrates consist of nitro-blue tetrazolium chloride (NBT) and 5-bromo-4-chloro-3'-indolyphosphate (BCIP), which, when reacted with alkaline phosphatase (alk-phos), form an intense, insoluble purple-black precipitate. This reaction provides a clear visual signal for detecting alkaline phosphatase activity on membranes or tissue samples.

Thermo Scientific™ offers these substrates in powder form as well as ready-to-use solutions for added convenience. The solution is designed to be sensitive and low-background, providing a reliable method for protein detection with alkaline phosphatase-conjugated antibodies.

Key Features

• AP Substrate: Specifically designed for the detection of alkaline phosphatase (alk-phos) activity, which is commonly used in Western blotting and immunohistochemistry (IHC) applications.
• Chromogenic Detection: The reaction produces a purple-black precipitate that is easily visible to the naked eye and can be photographed without requiring specialized equipment.
• Package Options: Available as individual powders for custom preparation, or as preformulated ready-to-use solutions. There are also options with or without a levamisole suppressor, which helps minimize endogenous phosphatase interference, ensuring more accurate results.

Highlights of Pierce NBT/BCIP 1-Step Solution (Part No. 34042)

• Best for Immunoblotting: The Pierce NBT/BCIP 1-Step Solution is ideal for immunoblotting applications, offering a sensitive and reliable method to detect alkaline phosphatase activity on nitrocellulose and PVDF membranes.
• Ready-to-Use: The solution is ready-to-use, reducing preparation time and potential for error.
• Low Background and High Sensitivity: It provides minimal assay-to-assay variability with low background, making it highly reliable for reproducible results.

Highlights of Pierce NBT/BCIP Plus Suppressor 1-Step Solution (Part No. 34070)

• Best for Immunohistochemistry: The Pierce NBT/BCIP Plus Suppressor Solution is specifically optimized for immunohistochemistry (IHC) applications. The inclusion of 1 mM levamisole inhibits endogenous phosphatase activity, reducing background staining and improving the clarity of target signals.
• Ready-to-Use: Like the standard version, it is a single component solution that is easy to use and does not require additional reagents or complex procedures.
• Sensitive and Clean Staining: Provides a dark purple precipitate in the presence of calf intestinal phosphatase while suppressing unwanted background staining.

Specifications

Applications

• Western Blotting: This substrate solution is ideal for detecting proteins in Western blotting experiments where alkaline phosphatase is used as the enzyme conjugate. The colorimetric reaction allows for easy visualization and documentation of the results.
• Immunohistochemistry (IHC): The solution is also highly suitable for IHC applications, where alkaline phosphatase is conjugated to antibodies for detecting specific antigens in tissue samples. The NBT/BCIP reaction produces a visible precipitate on tissue sections, enabling researchers to analyze the localization and expression of the target protein.
• In Situ Hybridization (ISH): The substrate can be used in ISH assays where alkaline phosphatase is used for the detection of specific nucleic acids within cells or tissues.

Storage and Handling

• Storage: Store the solution at 4°C upon receipt to maintain its stability and effectiveness.
• Shelf Life: The product has a long shelf life when stored at the recommended temperature. Always check the expiration date before use to ensure maximum performance.

Related Products

• 1-Step™ NBT/BCIP Plus Suppressor Substrate Solution
• NBT Substrate Powder (nitro-blue tetrazolium chloride)
• BCIP Substrate Powder (5-bromo-4-chloro-3'-indolyphosphate p-toluidine)

This product is for research use only and is not intended for diagnostic procedures.