Luminol assay for microdetermination of superoxide dismutase activity: its application to human fetal blood |
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Authors: | T P Huu C Marquetty C Pasquier J Hakim |
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Institution: | Laboratoire d''Immunologie et d''Hématologie, CHU X. Bichat, Université Paris 7, 16 rue Huchard, Paris 75018, France |
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Abstract: | A microtechnique for determining the superoxide dismutase activity in erythrocytes is described. This technique involves the inhibition of luminol-enhanced chemiluminescence of superoxide anion generated by xanthine-xanthine oxidase. Measurements required a steady-state chemiluminescence whether superoxide dismutase was present or absent; the level of luminescence was correlated to enzyme activity. Superoxide dismutase activity measured by this technique was 836 +/- 112 micrograms/g of hemoglobin for whole blood and 834 +/- 109 micrograms/g of hemoglobin for erythrocytes. When the reference technique was applied to larger amounts of blood, the results were 862 +/- 58 and 858 +/- 116 micrograms/g of hemoglobin for whole blood and washed erythrocytes, respectively. The enzymatic activity of superoxide dismutase from fetal blood (obtained by venipuncture in utero and of 19-26 weeks gestational age) was similar to that of adult blood, when measured by the new technique. |
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Keywords: | superoxide dismutase activity blood erythrocytes luminol chemiluminescence fetal blood |
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