Thermo Scientific™ NBT Substrate Powder (nitro-blue tetrazolium chloride), 1g

Thermo Scientific™ Pierce NBT Substrate Powder is a high-quality nitro-blue tetrazolium chloride (NBT) designed for use in chromogenic blotting and immunohistochemistry (IHC) applications. NBT is a tetrazolium salt that serves as an electron acceptor and undergoes reduction when exposed to alkaline phosphatase (alk-phos) activity. This reaction forms a dark blue-black formazan precipitate, making it ideal for detecting alkaline phosphatase in various biological samples.

NBT is commonly used in combination with 5-bromo-4-chloro-3'-indolyphosphate (BCIP) to visualize enzyme activity, especially for applications such as Western blotting, in situ hybridization, and immunohistochemistry. The product is available as a powder for flexibility in preparation, with the option to dissolve in water, buffers, or other solvents.

Key Features

• Alkaline Phosphatase Substrate: NBT is specifically designed for the detection of alkaline phosphatase (alk-phos) activity. It is commonly used in Western blotting, immunohistochemistry, and other assays requiring alkaline phosphatase-conjugated antibodies.
• Chromogenic Detection: NBT produces a purple-black precipitate upon reduction by alkaline phosphatase. This visible precipitate allows for easy observation without the need for specialized equipment, making it ideal for colorimetric detection.
• Membrane Compatibility: Compatible with nitrocellulose and PVDF membranes, which are commonly used in blotting techniques like Western blotting.
• Customizable: The substrate is available in powder form, providing flexibility for researchers who prefer to prepare custom solutions.

Highlights

• Sensitive Detection: The reaction produces an intense, insoluble formazan precipitate that is easily detectable by eye, making it suitable for a wide range of protein detection applications.
• Ready-to-Use Solutions Available: While this product is provided as a powder for flexible use, ready-to-use solutions are also available for those who prefer convenience. These solutions come with or without a levamisole suppressor to minimize background staining from endogenous phosphatases.
• Versatility: Ideal for applications requiring alkaline phosphatase detection, including immunoblotting and immunohistochemistry.

Specifications

Applications

• Western Blotting: Used to detect alkaline phosphatase-conjugated antibodies in protein samples. The resulting blue-black precipitate can be photographed or documented for further analysis.
• Immunohistochemistry (IHC): Suitable for detecting alkaline phosphatase activity in fixed tissue samples, allowing for the visual identification of target proteins.
• In Situ Hybridization: Used in in situ hybridization assays to detect nucleic acids, providing a visual signal for the presence of specific sequences.
• Northern and Southern Blotting: NBT can be used to detect nucleic acid sequences in Northern and Southern blotting experiments by visualizing alkaline phosphatase-linked probes.

Preparation Notes

• Solubility: NBT substrate powder is soluble in water, aqueous buffers, and dimethylformamide. A clear solution can be prepared at 20 mg/mL concentration for use in assays.
• Absorption: NBT has a maximum absorption at 260 nm in water. The reduced formazan product absorbs at 605 nm in nitrobenzole, providing a strong, visible color change.

Storage and Handling

• Storage: Store the substrate powder at 4°C and protect it from moisture. Ensure proper sealing of the container to maintain stability.
• Shipping: The product is shipped at ambient temperature, and it should be promptly stored at 4°C upon receipt to ensure optimal performance.

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)

Important Notes

• For Life Science Research Only: This product is intended for life science research purposes and is not for use in diagnostic procedures.

The Thermo Scientific™ NBT Substrate Powder is a valuable tool for researchers working with alkaline phosphatase detection, offering flexibility in preparation and reliable colorimetric results.