Rat leptin elisa

For F0 and F1 animals, one animal per litter underwent oral glucose tolerance testing (OGTT) (F0 n = 9, F1 n = 5–6). For F2 animals, two animals from each litter were tested (n = 8–14 from 4–7 litters). Following an overnight fast, at 09.00, blood was obtained by tail nicking into EDTA coated micro tubes (Starstedt, Germany) and plasma separated. 2 g/kg of 0.5 g/ml glucose solution (Sigma) was administered by oral gavage. Further blood was collected at 30 and 120 min.
For F0 animals, glucose was measured using a colorimetric kit (Cayman, USA) and for F1 and F2 with a kit (Alpha Laboratories Ltd., UK) adapted for use on a Cobas Fara centrifugal analyser (Roche, UK). Insulin was measured using a Rat Insulin ELISA kit (Mercodia, Sweden,). Fasting plasma triglyceride and cholesterol were determined using kits (Alpha Laboratories Ltd., UK and Olympus Diagnostics Ltd, UK, respectively), adapted for use on a Cobas Fara centrifugal analyser (Roche, UK). Terminal plasma leptin was measured using a Rat Leptin ELISA (Crystal-Chem, IL, USA).

Fasting glucose concentrations were measured in tail blood using a blood glucose analyser (AlphaTRAK, Abbott, Maidenhead, UK)). Fasting plasma insulin and leptin concentrations were determined by ELISA (ultrasensitive rat insulin EIA, Mercodia, Uppsala, Sweden; rat leptin ELISA, Crystal Chem, Downers Grove, IL, USA) and lipid content was measured using a Folch assay [15 (link)]. Lipid analysis was performed by the Core Biochemical Assay Laboratory (Cambridge, UK).

Fasting glucose concentrations were measured in tail blood using a blood glucose analyser (AlphaTRAK, Abbott, Maidenhead, UK)). Fasting plasma insulin and leptin concentrations were determined by ELISA (ultrasensitive rat insulin EIA, Mercodia, Uppsala, Sweden; rat leptin ELISA, Crystal Chem, Downers Grove, IL, USA) and lipid content was measured using a Folch assay [15 (link)]. Lipid analysis was performed by the Core Biochemical Assay Laboratory (Cambridge, UK).