Analysis Note

Absorption: NBT has its maximum absorption at 260 nm [in water],and the final product formazan has its maximum absorption at 605 nm [in nitrobenzole].

Application

Colony and plaque hybridizationImmunohistocytochemistryIn situ hybridizationNorthern blotSouthern blotWestern blot

NBT has been used in in situ hybridization.

General description

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. In immunolabeling technique, 4-Nitro blue tetrazolium (NBT) serves as an oxidant. The hydrolysis of 5-bromo-4-chloro-3-indolyl phosphate (BCIP) by alkaline phosphatase gives rise to leucoindigo, which is then being reduced by NBT to form an insoluble blue BCI. Certain cells reduce NBT, which is a colorless/yellow dye to blue or black formazan crystals. Due to this property, NBT is prefered in redox histochemistry and in biochemical applications. It is 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.

NBT is soluble in water, aqueous buffers, and dimethylformamide.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

Working solution: Solubility: soluble in water, aqueous buffers, and dimethylformamide. Can be dissolved in methanol. A clear solution is achieved at 20 mg/ml.

Quality

Performance-controlled in the LDH assay.