Mitochondria isolation from kidneys of Sham and CLP groups was performed using sucrose containing buffer as described previously [18 (link),19 (link)]. Mitochondrial complexes were extracted from the isolated mitochondria (250 μg) using 10% n-dodecyl-β-D-maltoside and 0.5 M aminocaproic acid (detergent/protein ratio, 2.5 g/g). The mitochondrial extracts (40 μg) were then resolved in a BN-PAGE gel [19 (link),20 (link)] followed by western blotting with 4-HNE (Abcam, 1:500) and Core-2 (Abcam,#ab14745;1:1000).
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Mitochondrial Complex Evaluation in Sepsis
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Other organizations : University of Arkansas for Medical Sciences
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Variable analysis
independent variables
- Sham group
- CLP group
- Mitochondrial complexes
- 4-HNE (oxidative stress marker)
- Core-2 (mitochondrial complex protein)
- Mitochondria isolation protocol
- Mitochondrial protein amount (250 μg) used for complex extraction
- Mitochondrial protein amount (40 μg) used for BN-PAGE and Western blotting
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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