Nitro blue tetrazolium chloride
{{chembox
| Watchedfields = changed
| verifiedrevid = 462261766
| ImageFile = Nitroblue_tetrazolium.svg
| ImageSize = 250px
| IUPACName = 2,2'-bis(4-Nitrophenyl)-5,5'-diphenyl-3,3'-(3,3'-dimethoxy-4,4'-diphenylene)ditetrazolium chloride
| OtherNames =
|Section1={{Chembox Identifiers
| ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}}
| ChemSpiderID = 8924
| 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
| InChIKey = FSVCQIDHPKZJSO-NUQVWONBAE
| StdInChI_Ref = {{stdinchicite|correct|chemspider}}
| StdInChI = 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
| StdInChIKey_Ref = {{stdinchicite|correct|chemspider}}
| StdInChIKey = FSVCQIDHPKZJSO-UHFFFAOYSA-L
| CASNo_Ref = {{cascite|correct|CAS}}
| CASNo = 298-83-9
| UNII_Ref = {{fdacite|correct|FDA}}
| UNII = X44P41F7ZK
| ChEMBL_Ref = {{ebicite|correct|EBI}}
| ChEMBL = 403063
| PubChem = 9281
| ChEBI_Ref = {{ebicite|correct|EBI}}
| ChEBI = 9505
| 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
}}
|Section2={{Chembox Properties
| Formula = C40H30Cl2N10O6
| MolarMass = 817.64 g/mol
| Appearance = yellow crystalline powder
| Density =
| MeltingPtC = 200
| BoilingPt =
| Solubility =
}}
|Section3={{Chembox Hazards
| MainHazards = may be reactive based on presence of tetrazole group, nitro group and contiguous nitrogen atoms
| FlashPt = not available
| AutoignitionPt =
| LD50 = 2 g/kg
}}
}}
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.{{cite journal |vauthors=Trinh le A, McCutchen MD, Bonner-Fraser M, Fraser SE, Bumm LA, McCauley DW |title=Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain |journal=BioTechniques |volume=42 |issue=6 |pages=756–9 |date=June 2007 |pmid=17612300 |doi=10.2144/000112476 |doi-access=free}}
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.{{cite journal |vauthors=Nisimoto Y, Wilson E, Heyl BL, Lambeth JD |title=NADH dehydrogenase from bovine neutrophil membranes. Purification and properties |journal=J. Biol. Chem. |volume=261 |issue=1 |pages=285–90 |date=5 January 1986|doi=10.1016/S0021-9258(17)42467-7 |pmid=3941077 |url=http://www.jbc.org/cgi/pmidlookup?view=long&pmid=3941077 |doi-access=free }}
Nitro blue tetrazolium is used in a diagnostic test,{{cite journal | last=Freeman | first=R |author2=King B | title=Technique for the performance of the nitro-blue tetrazolium (NBT) test | journal=Journal of Clinical Pathology |volume=25 | issue=10 | pages=912–914 |date=October 1972 |pmid=4119008 | doi=10.1136/jcp.25.10.912 | pmc=477548 }} particularly for chronic granulomatous disease and other diseases of phagocyte function. When there is an NADPH oxidase defect, the phagocyte is unable to make reactive oxygen species or radicals required for bacterial killing. As a result, bacteria may thrive within the phagocyte. The higher the blue score, the better the cell is at producing reactive oxygen species.{{cite journal |vauthors=Nathan DG, Baehner RL, Weaver DK |title=Failure of nitro blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease |journal=J. Clin. Invest. |volume=48 |issue=10 |pages=1895–904 |date=October 1969 |pmid=5387730 |pmc=322426 |doi=10.1172/JCI106156}}
References
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{{Myeloid blood tests}}
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{{DEFAULTSORT:Nitro Blue Tetrazolium Chloride}}
Category:Biochemistry detection reactions