Thermo Scientific™ Nitro blue tetrazolium chloride (NBT) ≥99%

Nitro Blue Tetrazolium Chloride (NBT) is a high-quality, colorless to yellow dye that is commonly used in cell biology and biochemical applications. As a tetrazolium salt, it has unique properties that allow it to be reduced into an insoluble blue formazan in certain cell types, making it useful for redox histochemistry. NBT is often used in combination with 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for chromogenic detection in immunolabeling and histochemistry techniques, particularly to visualize alkaline phosphatase activity.

NBT is highly soluble in water, aqueous buffers, and dimethylformamide, enabling easy preparation of working solutions for various assays. Its metabolic activity detection capability and ability to be reduced to blue or black formazan crystals make it a valuable tool in redox reactions and cell-based assays.

Key Features

• Solubility: NBT is soluble in water, aqueous buffers, and dimethylformamide, allowing for flexible use in a variety of experimental conditions. A clear solution can be prepared at a concentration of 20 mg/mL.
• Redox Histochemistry: NBT is widely used in redox histochemistry, where its ability to reduce to insoluble forazan crystals provides a visual color change. This reaction is especially valuable in distinguishing cellular activity or metabolic processes.
• Cellular Uptake: Due to its positive charge and tetrazole ring, NBT is taken up by cells via plasma membrane potential, which drives its use in cell biology to assess metabolic activity.
• Biochemical Applications: NBT is useful in immunolabeling techniques, such as Western blotting, Northern blotting, Southern blotting, and colony and plaque hybridization. It serves as an oxidizing agent for visualizing enzyme activity, especially alkaline phosphatase.

Applications

• Colony and Plaque Hybridization: Used for visualizing DNA/RNA hybridization in colony and plaque assays.
• Immunohistochemistry (IHC): NBT is used in immunohistochemistry to detect alkaline phosphatase activity in tissue samples.
• In Situ Hybridization: Effective in in situ hybridization for localizing specific nucleic acid sequences in tissue sections.
• Northern Blot and Southern Blot: Commonly used in Northern and Southern blotting to detect nucleic acid sequences after hybridization.
• Western Blot: Used in Western blotting as a colorimetric substrate for detecting alkaline phosphatase-conjugated antibodies.

Quality and Performance

• Performance-controlled in the LDH Assay: NBT is performance-controlled to ensure its consistency and reliability, particularly in the lactate dehydrogenase (LDH) assay, a widely used method for assessing cell viability and enzyme activity.

Preparation Notes

• Working Solution: NBT can be dissolved in water, aqueous buffers, dimethylformamide, or methanol. A clear solution can be achieved by dissolving at a concentration of 20 mg/mL.
• Absorption Spectra:
  • NBT has its maximum absorption at 260 nm in water.
  • The final forazan product (after reduction) absorbs at 605 nm in nitrobenzole.

Analysis Notes

• Absorption:
  • NBT absorbs maximally at 260 nm (in water).
  • The forazan product, the reduced form of NBT, absorbs maximally at 605 nm (in nitrobenzole), providing a clear signal for detecting enzyme activity.

Important Notes

• For Life Science Research Only: This product is intended for life science research purposes only and is not for diagnostic use.
• Not for Diagnostic Procedures: NBT is not to be used in any clinical diagnostic applications.

Specifications

NBT is an essential reagent for a wide range of research applications, particularly in cell biology and biochemical assays. Its redox activity and ability to generate a visible blue precipitate make it invaluable in assays requiring alkaline phosphatase detection.