Nitro blue tetrazolium chloride

Nitro blue tetrazolium chloride
Identifiers
CAS number	298-83-9 Y
PubChem	9281
ChemSpider	8924 Y
ChEBI	CHEBI:9505 Y
Jmol-3D images	Image 1
SMILES [Cl-].[Cl-].[O-][N+](=O)c1ccc(cc1)n2nc(n[n+]2c3ccc(cc3OC)c7ccc([n+]5nc(nn5c4ccc([N+]([O-])=O)cc4)c6ccccc6)c(OC)c7)c8ccccc8
InChI InChI=1S/C40H30N10O6.2ClH/c1-55-37-25-29(13-23-35(37)47-43-39(27-9-5-3-6-10-27)41-45(47)31-15-19-33(20-16-31)49(51)52)30-14-24-36(38(26-30)56-2)48-44-40(28-11-7-4-8-12-28)42-46(48)32-17-21-34(22-18-32)50(53)54;;/h3-26H,1-2H3;2*1H/q+2;;/p-2 Y Key: FSVCQIDHPKZJSO-UHFFFAOYSA-L Y InChI=1/C40H30N10O6.2ClH/c1-55-37-25-29(13-23-35(37)47-43-39(27-9-5-3-6-10-27)41-45(47)31-15-19-33(20-16-31)49(51)52)30-14-24-36(38(26-30)56-2)48-44-40(28-11-7-4-8-12-28)42-46(48)32-17-21-34(22-18-32)50(53)54;;/h3-26H,1-2H3;2*1H/q+2;;/p-2 Key: FSVCQIDHPKZJSO-NUQVWONBAE
Properties
Molecular formula	C40H30Cl2N10O6
Molar mass	817.64 g/mol
Appearance	yellow crystalline powder
Melting point	200°C
Hazards
Main hazards	may be reactive based on presence of tetrazole group, nitro group and contiguous nitrogen atoms
Flash point	not available
LD50	2gm/kg
Y blue tetrazolium chloride (verify) (what is: Y/N?) Except where noted otherwise, data are given for materials in their standard state (at 25 °C, 100 kPa)
Infobox references

Nitro blue tetrazolium is a chemical compound composed of two tetrazole moieties. It is used in immunology for sensitive detection of alkaline phosphatase (with BCIP). NBT serves as the oxidant and BCIP is the AP-substrate (and gives also dark blue dye).

Clinical significance

In immunohistochemistry the alkaline phosphatase is often used as a marker, conjugated to an antibody. The colored product can either be of the NBT/BCIP reaction reveals where the antibody is bound, or can be used in immunofluorescence.^[1]

The NBT/BCIP reaction is also used for colorimetric/spectrophotometric activity assays of oxidoreductases. One application is in activity stains in gel electrophoresis, such as with the mitochondrial electron transport chain complexes.^[2]

Nitro blue tetrazolium is used in a diagnostic test,^[3] particularly for chronic granulomatous disease and other diseases of phagocyte function. A disease where there is a defect in NADPH oxidase, therefore the phagocyte is unable to make the reactive oxygen species or radicals required for bacterial killing and results in bacteria thriving within the phagocyte. The higher the blue score, the better the cell is at producing reactive oxygen species.^[4]

References

1. ^ Trinh le A, McCutchen MD, Bonner-Fraser M, Fraser SE, Bumm LA, McCauley DW (June 2007). "Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain". BioTechniques 42 (6): 756–9. doi:10.2144/000112476. PMID 17612300. 
2. ^ Nisimoto Y, Wilson E, Heyl BL, Lambeth JD (5 January 1986). "NADH dehydrogenase from bovine neutrophil membranes. Purification and properties". J. Biol. Chem. 261 (1): 285–90. PMID 3941077. http://www.jbc.org/cgi/pmidlookup?view=long&pmid=3941077. 
3. ^ Freeman, R; King B (October 1972). "Technique for the performance of the nitro-blue tetrazolium (NBT) test". Journal of Clinical Pathology 25 (10): 912–914. doi:10.1136/jcp.25.10.912. PMC 477548. PMID 4119008. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=477548. 
4. ^ Nathan DG, Baehner RL, Weaver DK (October 1969). "Failure of nitro blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease". J. Clin. Invest. 48 (10): 1895–904. doi:10.1172/JCI106156. PMC 322426. PMID 5387730. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=322426.