Description
Nitro Blue Tetrazolium (NBT) is a chromogenic substrate that, like other tetrazolium compounds, can be reduced to produce a colored formazan derivative. It is used in the “NBT test” to evaluate the activity of NADPH oxidase in phagocytes, which results in the production of blue reduced NBT formazan in normal cells but not in those from patients with chronic granulomatous disease. NBT can also be used as a chromogenic activity stain for oxidoreductases in gels or solutions. More commonly NBT is often paired with 5-bromo-4-chloro-3-inolyl phosphate (PCIB) for the colorimetric detection of alkaline phosphatase activity. Alkaline phosphate converts PCIB to a product that reduces NBT to its formazan derivative, resulting in a black-purple precipitate.
Chemical Properties
NBT is a yellow crystals that can soluble in water, aqueous buffers, and dimethylformamide. Tetrazolium salts are useful in determining the metabolic activity of cells from many origins. The quaternary tetrazole ring core containing four nitrogen atoms, with a net positive charge induces cellular uptake through plasma membrane potential. This drives its use in cell biology.
Uses
Nitrotetrazolium Blue Chloride is a Tetrazolium dye used in colorimetric assays. It can be used to assess cell viability as a function of redox potential. Intense blue dye used in histochemistry determinations. actively respiring cells convert the water-soluble NBT to a deep blue-colored precipitate.
Application
Nitrotetrazolium Blue chloride has been used to detect the generation of oxygen metabolites in cells.
It has been used as a substrate for western blot detection.
It has been used for the staining of histological tissue sections.
Nitro Blue Tetrazolium chloride is used to confirm diagnosis of chronic granulomatous disease.
It is a microscopic stain, which is used to detect dehydrogenases and alkaline phosphatase. It acts as an NADPH-diaphorase substrate that competitively inhibits nitric oxide synthase.
Further, it is used in immunology for detection of alkaline phosphatase.
Definition
ChEBI: Nitro blue tetrazolium dichloride is an organic chloride salt having nitro blue tetrazolium(2+) as the counterion. It contains a nitro blue tetrazolium(2+).
General Description
Nitrotetrazolium Blue chloride (NBT) is a colorless to yellow dye that is reducible to blue or black formazan crystals by certain cells; formerly used to distinguish between nonbacterial and bacterial diseases, the latter causing neutrophils to reduce the dye; used to confirm diagnosis of chronic granulomatous disease.
Hazard
Moderately toxic by ingestion.
Biochem/physiol Actions
Nitrotetrazolium blue (NBT), in combination with BCIP (5-bromo-4-chloro-3-indolyl-phosphate), is required for the detection of labeled probes in western blots. It is part of the colorimetric detection assay. NBT and BCIP dyes detect alkaline phosphatase activity, thereby generating an insoluble purple dye (diformazan). The dye is visible at areas where the labeled probe has hybridized to its target.
Safety Profile
Moderately toxic by ingestion. When heated to decomposition it emits toxic vapors of NOx and Clí.
References
[1] R FREEMAN B K. Technique for the performance of the nitro-blue tetrazolium (NBT) test.[J]. Journal of Clinical Pathology, 1972, 25 10: 912-914. DOI:
10.1136/jcp.25.10.912[2] D G NATHAN D K W R L Baehner. Failure of nitro blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease.[J]. Journal of Clinical Investigation, 1969, 48 10: 1895-1904. DOI:
10.1172/jci106156[3] Y NISIMOTO. NADH dehydrogenase from bovine neutrophil membranes. Purification and properties.[J]. The Journal of Biological Chemistry, 1986, 261 1: 285-290.
[4] LE A TRINH. Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.[J]. BioTechniques, 2007, 42 6: 756-759. DOI:
10.2144/000112476
