Corticosterone elisa kit

On the last day of the experiment, the mice were fasted for 6 h (with free access to drinking water), and then blood and tissues were sampled. Blood samples were collected. The blood sample was centrifuged at 5000 rpm for 10 min to collect the serum, and the serum was frozen at -40°C until use. According to standard clinical protocols, the levels of serum glucose, low-density lipoprotein (LDL), high-density lipoprotein (HDL), total cholesterol, and triglyceride (TG) were measured using Hitachi 7600 biochemical analyzer (Hitachi, Japan). In addition, serum insulin and corticosterone levels were evaluated using the ultrasensitive mouse insulin ELISA kit (CrystalChem, IL, USA) and the commercially available corticosterone ELISA kit (Abnova, Taiwan, China) according to the manufacturers’ instructions.

We performed ELISA to measure the concentration of corticosterone and tumor necrosis factor alpha (TNF-alpha) in sera of animals; the corresponding assay systems were purchased from Abnova (Corticosterone ELISA Kit, Abnova GmbH, Heidleberg, Germany, cat. # B0AP01090J00015) and Thermo Fischer Scientific (Rat TNF-alpha ELISA Kit, cat. # ER3TNFA), respectively. Samples were assayed in duplicates adhering strictly to manufacturer directions. Optical density of samples on the 96 well ELISA plate was measured using Spectramax M2 (Molecular Devices, Sunnyvale, CA, USA) and computed with SoftMax Pro, V5 software (Molecular Devices).

Serum concentrations of corticosterone were determined using the Corticosterone ELISA kit (Abnova). Samples were diluted at 1:50 and the assay was performed according to the manufacturer’s instructions. Serum concentrations of IGF-1 and IGFBP-3 were determined using the Mouse/Rat IGF-1 Quantikine ELISA kit (R&D Systems) and IGFBP-3 DuoSet ELISA kit (R&D Systems), respectively. Samples were diluted at 1:500 and the assay was performed according to the manufacturer’s instructions. Serum concentrations of total IgE were quantified with BD OptEIA™ Mouse IgE ELISA Set (BD Biosciences, San Chose, Canada) with samples diluted 1:100 according to the manufacturer’s instructions. Samples were assayed in monoplicate.