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Leptin, a hormone produced by fat cells that regulates appetite and energy expenditure, decreases during weight loss

After centrifugation (1200 g , 4 C, 10 min) 9 mL acidified H 2 O was added (final pH = 3.5) and samples were subjected to solid phase extraction

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Total GLP-1 was analyzed by a chemiluminescent enzyme-linked assay (Mercodia, Uppsala, Sweden, Catalog # 10-1278-01, RRID: AB_2892202) with, according to the manufacturer, a 100% specificity to the GLP-1 (936) amide

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abdomen, thigh, or upper arm sites