A highly sensitive method for determining both Mn- and Cu-Zn superoxide dismutase activities in tissues and blood cells.
A highly sensitive method for determining the superoxide dismutase (EC 1.15.1.1) in various tissues and blood cells is described. This method involves inhibition of a cypridina luciferin analog that is chemiluminescence dependent upon O2- generated by hypoxanthine-xanthine oxidase. Manganeous superoxide dismutase, which is sensitive to sodium dodecyl sulfate, was determined and calculated by subtraction of superoxide dismutase activity in tissue extract treated with this detergent (Cu-Zn superoxide dismutase) from that in untreated tissue extract (total superoxide dismutase). Both Mn- and Cu-Zn superoxide dismutase activities were expressed as equivalent nanograms of bovine erythrocyte superoxide dismutase per milliliter. Sensitivity limits of the chemiluminescence methods with 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-alpha]pyraz in-3-one and 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2-alpha]pyrazin-3-one as cypridina luciferin analogs were 1 ng and 2-3 ng of superoxide dismutase/ml, respectively.
College of Medical Care and Technology, Gunma University, Japan.
M
M
Nakano
H
H
Kimura
M
M
Hara
M
M
Kuroiwa
M
M
Kato
K
K
Totsune
T
T
Yoshikawa
D000818:Animals; D004912:Erythrocytes / Q000201:enzymology*; D007093:Imidazoles; D008163:Luminescent Measurements; D008297:Male; D009928:Organ Specificity; D011719:Pyrazines; D051381:Rats; D011919:Rats, Inbred Strains; D012680:Sensitivity and Specificity; D013482:Superoxide Dismutase / Q000097:blood*
D007093:Imidazoles; D011719:Pyrazines; C063412:2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one; D013482:Superoxide Dismutase