Sensitive spectrophotometric method for the determination of superoxide dismutase activity in tissue extracts
Journal Article
·
· Anal. Biochem.; (United States)
Superoxide dismutase (EC 1.15.1.1) has been assayed by a spectrophotometric method based on the inhibition of a superoxide-driven NADH oxidation. The assay consists of a purely chemical reaction sequence which involves EDTA. Mn(II), mercaptoethanol, and molecular oxygen, requiring neither auxiliary enzymes nor sophisticated equipment. The method is very flexible and rapid and is applicable with high sensitivity to the determination of both pure and crude superoxide dismutase preparations. The decrease of the rate of NADH oxidation is a function of enzyme concentration, and saturation levels are attainable. Fifty percent inhibition, corresponding to one unit of the enzyme, is produced by approximately 15 ng of pure superoxide dismutase. Experiments on rat liver cytosol have shown the specificity of the method for superoxide dismutase. Moreover, common cellular components do not interfere with the measurement, except for hemoglobin when present at relatively high concentrations. The assay is performed at physiological pH and is unaffected by catalase.
- Research Organization:
- Universita di Firenze, Italy
- OSTI ID:
- 5533630
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 154:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400104* -- Spectral Procedures-- (-1987)
550200 -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BODY
CARBOXYLIC ACIDS
CHELATING AGENTS
CHEMICAL ANALYSIS
CHEMICAL REACTIONS
COENZYMES
DIGESTIVE SYSTEM
EDTA
ELEMENTS
ENZYME ACTIVITY
ENZYME INHIBITORS
ENZYMES
GLANDS
GLOBIN
HEMOGLOBIN
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INTERFERING ELEMENTS
KINETICS
LIVER
MAMMALS
MANGANESE
MATERIALS
METALS
NADH2
NONMETALS
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
OXIDATION
OXIDOREDUCTASES
OXYGEN
PH VALUE
PIGMENTS
PORPHYRINS
PROTEINS
QUANTITATIVE CHEMICAL ANALYSIS
RATS
REACTION KINETICS
RODENTS
SENSITIVITY
SPECIFICITY
SPECTROPHOTOMETRY
SUPEROXIDE DISMUTASE
THIOLS
TISSUE EXTRACTS
TRANSITION ELEMENTS
VERTEBRATES
400104* -- Spectral Procedures-- (-1987)
550200 -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BODY
CARBOXYLIC ACIDS
CHELATING AGENTS
CHEMICAL ANALYSIS
CHEMICAL REACTIONS
COENZYMES
DIGESTIVE SYSTEM
EDTA
ELEMENTS
ENZYME ACTIVITY
ENZYME INHIBITORS
ENZYMES
GLANDS
GLOBIN
HEMOGLOBIN
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INTERFERING ELEMENTS
KINETICS
LIVER
MAMMALS
MANGANESE
MATERIALS
METALS
NADH2
NONMETALS
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
OXIDATION
OXIDOREDUCTASES
OXYGEN
PH VALUE
PIGMENTS
PORPHYRINS
PROTEINS
QUANTITATIVE CHEMICAL ANALYSIS
RATS
REACTION KINETICS
RODENTS
SENSITIVITY
SPECIFICITY
SPECTROPHOTOMETRY
SUPEROXIDE DISMUTASE
THIOLS
TISSUE EXTRACTS
TRANSITION ELEMENTS
VERTEBRATES