Nitro blue tetrazolium chloride: Difference between revisions
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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]].<ref>{{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}}</ref> | 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]].<ref>{{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}}</ref> | ||
The NBT/BCIP reaction is also used for colorimetric/[[spectrophotometry|spectrophotometric]] [[enzyme assay|activity assays]] of [[oxidoreductase]]s. One application is in [[activity stain]]s in [[gel electrophoresis]], such as with the mitochondrial [[electron transport chain]] complexes.<ref>{{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 | | The NBT/BCIP reaction is also used for colorimetric/[[spectrophotometry|spectrophotometric]] [[enzyme assay|activity assays]] of [[oxidoreductase]]s. One application is in [[activity stain]]s in [[gel electrophoresis]], such as with the mitochondrial [[electron transport chain]] complexes.<ref>{{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 |doi-access=free }}</ref> | ||
Nitro blue tetrazolium is used in a diagnostic test,<ref>{{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 }}</ref> 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.<ref>{{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}}</ref> | Nitro blue tetrazolium is used in a diagnostic test,<ref>{{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 }}</ref> 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.<ref>{{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}}</ref> | ||
Latest revision as of 09:00, 28 June 2025
Template:Chembox 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. 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.[4]