Left ventricular tissue was homogenized in PBS on ice and centrifuged at 4°C, at 10,000 rpm for 10 min. The supernatant was collected for measurements. As descripted in the manufacturer’s protocol and in our studies (El-Mas et al., 2012 (link), Ibrahim et al., 2014 (link), Yao and Abdel-Rahman, 2016 (link)), 5 μg of sample protein was used to measure the catalytic activity by colorimetric catalase assay kit (Sigma-Aldrich, St. Louis, MO). The absorbance was read at 520 nm. A mitochondrial ALDH2 activity assay kit (Abcam, Ann Arbor, MI) was used for the measurement of mitALDH2 activity in 500 μg sample protein; the absorbance was read at 450 nm every 3 min for 1 hr.
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Enzymatic Activity Assay for Left Ventricular Tissue
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Corresponding Organization : East Carolina University
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Variable analysis
independent variables
- Homogenization of left ventricular tissue in PBS on ice
- Centrifugation at 4°C, 10,000 rpm for 10 min
- Catalytic activity of catalase measured by colorimetric assay (absorbance at 520 nm)
- Mitochondrial ALDH2 activity measured by assay kit (absorbance at 450 nm every 3 min for 1 hr)
- Temperature at 4°C during centrifugation
- Protein amount used for catalase activity (5 μg) and mitALDH2 activity (500 μg) measurements
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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