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MB Sample ID: SA057698
Local Sample ID: | 10_25_2016_islet_OC_14 |
Subject ID: | SU001003 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL/6N |
Age Or Age Range: | 4 weeks old (young) or 10-15 months old (old) |
Gender: | Male and female |
Animal Animal Supplier: | Charles River Labs |
Animal Feed: | standard chow diet (PicoLab Rodent Diet 20 pellets, #5053 |
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Treatment:
Treatment ID: | TR001017 |
Treatment Summary: | Islets were starved for 1 hour in KRBH supplemented with 2.8 mM glucose as for GSIS assays. Islets were then transferred to petri dishes containing U-13C-labeled glucose as indicated and immediately seeded into wells of non-treated 12-well plates at a density of 100 size-matched islets per replicate in a final volume of 500 μl KRBH containing basal (2.8 mM) or stimulatory (16.8 mM) concentrations of labeled glucose. Tracing was performed for the indicated durations at 37oC with 5% CO2. Following the trace, islets were rapidly chilled by swirling plate to resuspend islets then transferring entire contents of each well to an Eppendorf tube on ice. Islets were then pelleted by centrifugation at 500 x g for 2 min at 4oC, washed in 1 mL of ice cold 0.9% NaCl, centrifuged as before, then pellets were frozen in a dry ice/ethanol bath and stored at -80oC. |