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MB Sample ID: SA018721
Local Sample ID: | 061215byusa1099_1 |
Subject ID: | SU000414 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57Bl/6J |
Age Or Age Range: | 4 months |
Gender: | Female |
Human Smoking Status: | Former/Current |
Animal Animal Supplier: | Jackson Laboratories |
Animal Housing: | housed in a facility at 23°C |
Animal Light Cycle: | 12∶12 hr light cycle, lights on 0700 |
Animal Feed: | with free access to chow (4.5% fat/weight), for at least 1 week before being studied |
Species Group: | Mammal |
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Sample Preparation:
Sampleprep ID: | SP000421 |
Sampleprep Summary: | Tissue was homogenized using a glass/Teflon Potter-Elvehjem tissue grinder (240 rpm) and fractionated by centrifugation at 800 g (10 min), and the supernatant collected and spun at 12000 g (9 min). The pellet was resuspended in 20 ml BM and incubated on ice for 5 min (myofibrillar repolymerization). Samples were spun at 800 g (8 min) to pellet actin-myosin polymers. The supernatant was then spun at 12000 g (9 min). The final pellet was resuspended in 220 µl of BM. |
Sampleprep Protocol Filename: | Long-Chain_Fatty_Acid_Metabolite_Profiles_in_Skeletal_Muscle_Mitochondria.PDF |
Sampleprep Protocol Comments: | This isolation procedure yields mitochondria with high respiratory control ratios (state 3/state 4; ∼8–10 when supplied with 10 mM pyruvate/5 mM malate), and which are capable of activating palmitate [29], a process dependent on the integrity of enzymes on the mitochondrial outer membrane. Protein concentration was determined by a modified Lowry method with BSA as standard. |
Processing Method: | Homogenized |