Testicular tissues (100 mg) were washed with 1X PBS (pH 7.4) to remove excess blood thoroughly. The activity of SOD, GPx, and catalase in whole tissue supernatant was evaluated using commercially available kits (Cayman Chemical, Ann Arbor, MI, USA; item no.706002, superoxide dismutase kit; item no. 703102, glutathione peroxidase kit; item no. 707002, catalase assay kit) as per manufacturer’s instructions. Values were expressed as milligrams of protein.
Glutathione peroxidase kit
Manufactured by Cayman Chemical
Sourced in United States
The Glutathione peroxidase kit is a laboratory tool used to measure the activity of the enzyme Glutathione peroxidase. Glutathione peroxidase is an important antioxidant enzyme that helps protect cells from oxidative stress by catalyzing the reduction of hydrogen peroxide and other organic hydroperoxides.
Automatically generated - may contain errors
7 protocols using glutathione peroxidase kit
1
Antioxidant Enzyme Activities in Testis
2
Measuring Muscle GPx Activity
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The GPx activity in muscle homogenates was determined by Glutathione Peroxidase Kit (703102, Cayman Chemicals, USA) in accordance with supplied manufacturer's instruction.
The activity of GPx was expressed as nanomoles per minute per milligram of protein.
The activity of GPx was expressed as nanomoles per minute per milligram of protein.
3
Antioxidant Enzyme Activity Assays
4
Cerebral GPx Enzymatic Activity Assay
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A Glutathione Peroxidase Kit (Item No. 703102) was used to measure GPx enzymatic activity following the manufacturer's instructions (Cayman Chemical Company; Ann Arbor, MI, USA). After perfusion with PBS, cerebral tissues were homogenized with 200 μL of cold GPx sample buffer 10X (50 mM Tris-HCl, pH 7.5, 5 mM EDTA, and 1 mM DTT). In a 96-well plate, the background control wells contained 70 μL of a GPx assay buffer (10X), 50 μL of the cosubstrate mixture, and 50 μL of NADPH. Positive control wells were supplemented with 50 μL of an assay buffer, 50 μL of a cosubstrate mixture, 50 μL of NADPH, and 20 μL of a diluted GPx Control. Subsequently, the background and positive control wells were read at 340 nm in triplicate, recording the NADPH absorbance change during each minute for 5 min. The sample wells contained 70 μL of an assay buffer, 50 μL of the cosubstrate mixture, 50 μL of NADPH, and 20 μL of a supernatant sample. The reaction was initiated by adding 20 μL of cumene hydroperoxide to all wells. Finally, the absorbance at 340 nm was recorded every minute for 5 min using a microplate reader (Bechmark™, Bio-Rad; Hercules, California 94547, USA). The results of enzymatic activity were reported as NADPH nmol/min−1/mL.
5
Glutathione Peroxidase Activity Assay
6
Antioxidant Enzyme Analysis in Testis
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Left testis tissues (100 mg) were rinsed with 1X PBS (pH 7.4) to remove excess blood thoroughly. Activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase in whole tissue supernatants were evaluated using commercial kits (item No. 706002, superoxide dismutase kit; item No. 703102, glutathione peroxidase kit; item no. 707002, catalase assay kit, Cayman Chemical, Ann Arbor, MI, USA) per the manufacturer’s instructions. Antioxidant enzymes’ activities are expressed as per milligram of protein.
7
Antioxidant Enzyme Quantification in Testis
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Testis tissues (100 mg) were rinsed with 1X PBS (pH 7.4) to remove any red blood cells and clots. Concentrations of SOD, GPx, and catalase in whole tissue supernatant were measured using commercial kits (item no. 706002, superoxide dismutase kit; item no. 703102, glutathione peroxidase kit; item no. 707002, catalase assay kit, Cayman Chemical, Ann Arbor, MI, USA) following the manufacturer’s instructions. Values are expressed as per milligram protein.
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