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Superoxide dismutase (SOD) assay

MJ Malsawmhriatzuala Jeremy
GG Guruswami Gurusubramanian
VR Vikas Kumar Roy
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The previously described method was adopted for the superoxide dismutase assay73. Briefly, a mixture containing 50 µl of 10% tissue homogenate, 600 µl of 52 mM sodium pyrophosphate buffer (pH 8.3), 50 µl of 186 µM Phenazine methosulphate (PMS), 150 µl of 300 µM Nitroblue tetrazolium(NBT). The reaction was started by the addition of 100 µl of 750 µM ml NADH and incubated at 30 °C. The reaction was terminated after 90 seconds incubation by the addition of 500 µl glacial acetic acid. Two ml of n-butanol was added, vortexed and allowed to stand for 10 minutes. The mixture was centrifuged at 10000 × g for 10 minutes at room temperature and the supernatant was collected. Color intensity was measured at 560 nm and the concentration of SOD was expressed as units (U)/mg protein. A mixture devoid of tissue homogenate was taken as blank.

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