Hippocampal catalase activity was analyzed as previously described [30 (link)]. The hippocampal homogenates were mixed with phosphate buffer (250 mM, pH 7.0), methanol (12 mM), and H2O2 (44 mM) in sequence. After incubation at RT for 20 min, the reaction was stopped by the addition of Purpald solution (22.8 mM Purpald in 2 N potassium hydroxide). The mixture was incubated at 25 °C for 20 min; then, potassium periodate (65.2 mM in 0.5 N potassium hydrate) was added. Absorbance at 550 nm was measured using a UV spectrophotometer, and the results were calculated using a catalase standard curve.
Bovine erythrocyte sod
Bovine erythrocyte SOD is a laboratory product that contains the enzyme superoxide dismutase (SOD) isolated from bovine erythrocytes. SOD is an antioxidant enzyme that catalyzes the conversion of superoxide radicals into oxygen and hydrogen peroxide.
6 protocols using bovine erythrocyte sod
Oxidative Stress Assays in Hippocampus
Hippocampal catalase activity was analyzed as previously described [30 (link)]. The hippocampal homogenates were mixed with phosphate buffer (250 mM, pH 7.0), methanol (12 mM), and H2O2 (44 mM) in sequence. After incubation at RT for 20 min, the reaction was stopped by the addition of Purpald solution (22.8 mM Purpald in 2 N potassium hydroxide). The mixture was incubated at 25 °C for 20 min; then, potassium periodate (65.2 mM in 0.5 N potassium hydrate) was added. Absorbance at 550 nm was measured using a UV spectrophotometer, and the results were calculated using a catalase standard curve.
Measurement of Hepatic Antioxidant Enzymes
CAT activity in the hepatic tissue was determined using the method of the previously described method [26 (link)]. Briefly, 140 μL of phosphate buffer (250 mM, pH 7.0), 150 μL of 12 mM methanol, and 30 μL of H2O2 were mixed with 300 μL of standard or diluted hepatic tissue homogenate in a 13 × 100 mm test tube. The reaction was allowed to proceed for 10 to 20 min and was stopped by the addition of 450 μL of Purpald (22.8 mM in 2 N potassium hydroxide). The mixture was maintained for 20 min at 25°C, and then 150 μL of potassium periodate (65.2 mM in 0.5 N potassium hydrate) was added to the same tube. The absorbance was measured at 550 nm using a microplate reader (Molecular Devices).
Hepatic Antioxidant Enzyme Assays
Catalase activity in the hepatic tissue was determined according to the previous method [26] (link). The protein sample reacted with hydrogen and Purpald solution (22.8mM of Purpald in 2N potassium hydroxide). The absorbance at 550 nm of the purple formaldehyde adduct was measured using a spectrophotometer.
Enzymatic Activities Measurement Protocol
Catalase activity was determined as previously described [20 (link)]. The absorbance of the purple formaldehyde adduct was measured at 550 nm using a spectrophotometer (Molecular Devices Corp.).
Hippocampal Oxidative Stress and Antioxidants
Cerebral Cortex and Hippocampus Catalase and SOD Assays
SOD activity in cerebral cortex and hippocampus was determined using a SOD assay kit (Dojindo Laboratories, Kumamoto, Japan), according to the protocol from the manufacture. Bovine erythrocyte SOD (Sigma) was used as a standard (0.01 - 20 U/mL).
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