TY - JOUR
T1 - Electrophoretic detection of ascorbate oxidase activity by photoreduction of nitroblue tetrazolium
AU - Maccarrone, Mauro
AU - Rossi, Antonello
AU - D'Andrea, Gabriele
AU - Amicosante, Gianfranco
AU - Avigliano, Luciana
PY - 1990
Y1 - 1990
N2 - A method for the detection of ascorbate oxidase in electrophoretic gels is described. This method relies on the ability of the enzyme to prevent the photoreduction of nitroblue tetrazolium (NBT). The method is based on that described by C. Beauchamp and I. Fridovich (1971, Anal. Biochem. 44, 276-287) for the superoxide dismutase and was made specific for ascorbate oxidase detection by treating the gel with 0.1 m hydrogen peroxide. Ascorbate (25 μm) or riboflavin (500 μm) was used as the electron donor. The possible reaction mechanism in the presence of ascorbate has been investigated. Western and Northern blot analyses confirmed the results obtained from the NBT staining procedure.
AB - A method for the detection of ascorbate oxidase in electrophoretic gels is described. This method relies on the ability of the enzyme to prevent the photoreduction of nitroblue tetrazolium (NBT). The method is based on that described by C. Beauchamp and I. Fridovich (1971, Anal. Biochem. 44, 276-287) for the superoxide dismutase and was made specific for ascorbate oxidase detection by treating the gel with 0.1 m hydrogen peroxide. Ascorbate (25 μm) or riboflavin (500 μm) was used as the electron donor. The possible reaction mechanism in the presence of ascorbate has been investigated. Western and Northern blot analyses confirmed the results obtained from the NBT staining procedure.
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U2 - 10.1016/0003-2697(90)90534-G
DO - 10.1016/0003-2697(90)90534-G
M3 - Article
C2 - 2221347
AN - SCOPUS:0025324873
VL - 188
SP - 101
EP - 104
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -