Ab193687

Blood samples were collected from the anterior cephalic vein after the dogs were subjected to fasting for at least 16 h at the beginning (0 weeks) and the end (16 weeks) of the experiment. The collected blood was immediately divided between EDTA vacutainer tubes (ref 367861, BD Vacutainer, NJ, USA) and serum vacutainer tubes (ref 367812, BD Vacutainer). The blood collected in the EDTA tubes were used in the complete blood cell count (CBC) analysis using an automated blood analyzer ProCyte Dx (IDEXX Laboratories, Westbrook, ME, USA) immediately after collection. The blood collected in the serum vacutainer tubes were centrifuged at 1,650×g for 15 min to separate the serum; the serum supernatant was stored at –80°C until further use. Serum biochemical composition analysis was performed using an automated biochemistry analyzer, Hitachi 7180 (Hitachi High-Technologies, Tokyo, Japan). Adiponectin (MBS2607889, MyBioSource, San Diego, CA, USA), immunoglobulin G (IgG, ab157701, Abcam, Cambridge, MA, USA), interleukin-6 (IL-6, ab193686, Abcam), insulin (MBS031096, MyBioSource), leptin (MBS037935, MyBioSource), and tumor necrosis factor-α (TNF-α, ab193687, Abcam) were measured by enzyme-linked immunosorbent assay using a microplate spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) in duplicates, according to the manufacturer’s instructions.