Levels of pro-inflammatory IL-6 and leptin were determined in plasma, by using a Mouse IL-6 ELISA kit (eBioscience®, ThermoFisher Scientific, Illkirch Cedex, France) and a Mouse Leptin ELISA kit (RayBiotech®, Norcross, GA, USA), respectively. The assays were performed according to the manufacturer’s instructions and the absorbance was measured at 450 nm (FLUOstar Optima®, Bmg Labtech, Offenburg, Germany).
Mouse leptin elisa kit
Manufactured by RayBiotech
Sourced in France, United States
The Mouse Leptin ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay designed for the measurement of mouse leptin levels in serum, plasma, and cell culture supernatants. The kit utilizes a specific antibody coated on a 96-well plate to capture leptin, and a detection antibody is used to quantify the captured leptin.
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3 protocols using mouse leptin elisa kit
1
Quantifying Plasma IL-6 and Leptin
2
Measuring Serum Leptin and CXCL2 Levels
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The leptin concentration in serum was measured using a Mouse Leptin ELISA Kit (RayBiotech, Norcross, GA, USA) or a Human Leptin ELISA Kit (NeoBioscience Technology Co., Ltd, China). The CXCL2 concentration in the cell supernatant was measured using a Mouse CXCL2 ELISA Kit (RENJIEBIO Co, Ltd, China). The assays were carried out according to the manufacturer’s instructions.
3
Comprehensive Body Composition Analysis
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To compile growth curves, we weighed the mice once per week for 25 weeks, from weaning. Magnetic resonance imaging and nuclear magnetic resonance were used to measure body composition (fat and lean mass) in mice. At the end of the experiments, the mice were sacrificed, and their liver; kidneys; BAT; and various fat pads, including perirenal, gonadal, and inguinal WAT, were dissected and weighed. As a marker of body fat mass, we measured circulating leptin. To achieve this objective, we obtained plasma by using 0.5 M EDTA (25 μl) and centrifugation from blood acquired during the sacrifice of the mice. Leptin concentration was measured by using a mouse leptin ELISA kit (RayBiotech). To measure food intake, we housed the mice in individual cages. After 3 days of acclimation to individual housing, daily and cumulative food intake were measured over a 4-day period.
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