Source 1review1997Journal of Biological Chemistry
Toolkit/nitroblue tetrazolium superoxide assay
nitroblue tetrazolium superoxide assay
Also known as: nitroblue tetrazolium
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
One additional artifactual detector of O·̄2 needs to be mentioned because of its widespread misuse, and that is nitroblue tetrazolium... many aerobic tetrazolium reductions are inhibitable by SOD even though O·̄2 was not being produced in the system in the absence of the tetrazolium.
Usefulness & Problems
Why this is useful
Nitroblue tetrazolium is used as a detector through reduction to formazan products. The review argues that radical intermediates formed during this process can themselves generate superoxide.; colorimetric detection of redox activity often interpreted as superoxide-related
Source:
Nitroblue tetrazolium is used as a detector through reduction to formazan products. The review argues that radical intermediates formed during this process can themselves generate superoxide.
Source:
colorimetric detection of redox activity often interpreted as superoxide-related
Problem solved
It is used to report redox activity in systems where superoxide is of interest.; attempts to provide a detector for superoxide
Source:
It is used to report redox activity in systems where superoxide is of interest.
Source:
attempts to provide a detector for superoxide
Problem links
attempts to provide a detector for superoxide
LiteratureIt is used to report redox activity in systems where superoxide is of interest.
Source:
It is used to report redox activity in systems where superoxide is of interest.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Techniques
Functional AssayTarget processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The assay requires nitroblue tetrazolium and a readout of formazan formation, but the reagent chemistry introduces self-generated signal artifacts.; interpretation is confounded by assay-generated superoxide
It does not cleanly distinguish pre-existing superoxide production from superoxide generated by the detector chemistry itself.; artifactual detector of superoxide; tetrazolium radical intermediates can reduce oxygen to superoxide; SOD inhibition can occur even when superoxide was not originally being produced
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Detector molecules such as ferricytochrome c and spin-trapping agents are used to detect superoxide, but they are not specific for superoxide and often rely on SOD inhibition to lend specificity.
Lucigenin luminescence is an inappropriate specific detector of superoxide because lucigenin chemistry can itself generate superoxide and increase intracellular superoxide production.
Luminol luminescence is misused as a superoxide measurement because the luminol radical can generate superoxide and the signal can be caused by multiple oxidants.
Nitroblue tetrazolium is an artifactual superoxide detector because tetrazolium radical intermediates can generate superoxide, making SOD-inhibitable signal possible even when superoxide was not initially present.
Rapid inactivation of [4Fe-4S]-containing dehydratases such as aconitase can be used as a reliable measure of intracellular superoxide, although other oxidants such as peroxynitrite can also inactivate aconitase.
Approval Evidence
1 source1 linked approval claimfirst-pass slug nitroblue-tetrazolium-superoxide-assay
One additional artifactual detector of O·̄2 needs to be mentioned because of its widespread misuse, and that is nitroblue tetrazolium... many aerobic tetrazolium reductions are inhibitable by SOD even though O·̄2 was not being produced in the system in the absence of the tetrazolium.
Source:
review summarysupports
Nitroblue tetrazolium is an artifactual superoxide detector because tetrazolium radical intermediates can generate superoxide, making SOD-inhibitable signal possible even when superoxide was not initially present.
Source:
Comparisons
Source-stated alternatives
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Source:
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Source-backed strengths
widely used detector format
Source:
widely used detector format
Compared with ferricytochrome c superoxide detection assay
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Shared frame: source-stated alternative in extracted literature
Strengths here: widely used detector format.
Relative tradeoffs: artifactual detector of superoxide; tetrazolium radical intermediates can reduce oxygen to superoxide; SOD inhibition can occur even when superoxide was not originally being produced.
Source:
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Compared with lucigenin luminescence assay
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Shared frame: source-stated alternative in extracted literature
Strengths here: widely used detector format.
Relative tradeoffs: artifactual detector of superoxide; tetrazolium radical intermediates can reduce oxygen to superoxide; SOD inhibition can occur even when superoxide was not originally being produced.
Source:
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Compared with luminol luminescence assay
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Shared frame: source-stated alternative in extracted literature
Strengths here: widely used detector format.
Relative tradeoffs: artifactual detector of superoxide; tetrazolium radical intermediates can reduce oxygen to superoxide; SOD inhibition can occur even when superoxide was not originally being produced.
Source:
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Compared with spin-trapping superoxide detection
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Shared frame: source-stated alternative in extracted literature
Strengths here: widely used detector format.
Relative tradeoffs: artifactual detector of superoxide; tetrazolium radical intermediates can reduce oxygen to superoxide; SOD inhibition can occur even when superoxide was not originally being produced.
Source:
The review contrasts nitroblue tetrazolium with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and aconitase-based measurement.
Ranked Citations
- 1.