Igf 1 elisa kit

After 8 weeks of treatment, blood samples were obtained from the mice by cardiac puncture at the time of sacrifice. The samples were centrifuged at 800× g for 15 min at 4 °C, and the serum samples obtained were stored at −80 °C until subsequent analysis. The serum concentrations of IL-6, IL-1β, and TNF-α were measured using a Mouse High-sensitivity T Cell Magnetic Bead Panel (Merck Millipore, Burlington, MA, USA). The serum concentrations of estradiol and IGF-1 were measured using a Mouse Estradiol ELISA Kit (MyBioSource, San Diego, CA, USA) and an IGF-1 ELISA kit (Thermo Fisher Scientific, Waltham, MA, USA), respectively. All these analyses were performed according to the manufacturers’ instructions. Absorbances were measured at appropriate wavelengths using a Luminex 100 analyzer (Luminex, Austin, TX, USA). All the sample concentrations were measured in triplicate.

Blood samples were collected by cardiac puncture at the time of sacrifice and centrifuged at 800× g for 15 min at 4 °C, and the serum samples obtained were stored at −80 °C. The serum concentrations of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were measured using a mouse Th17 magnetic bead panel (Merck Millipore, MA, USA). The serum concentration of insulin-like growth factor (IGF)-1 was measured using an IGF-1 ELISA kit (Thermo Fisher Scientific, Waltham, MA, USA). The serum concentration of glutathione was measured using a Glutathione assay kit (Cayman chemical, Ann Arbor, MI, USA). All these analyses were performed according to the manufacturers’ instructions. Absorbances were measured at appropriate wavelengths using a Luminex 100 analyzer (Luminex, Austin, TX, USA). All measurements were made in triplicate.

The following plasma metabolites were measured: glucose, urea, uric acid, creatinine, bilirubin, and the hepatic enzymes glutamic oxaloacetic transaminase (GOT), glutamate pyruvate transaminase (GPT) and gamma-glutamyl transferase (GGT). These metabolites were analyzed using commercial kits, according to the manufacturer’s instructions, and a Hitachi 737 Automatic Analyser (Hitachi Ltd., Tokyo, Japan). The plasma levels of cytokines were determined by Enzyme-Linked ImmunoSorbent Assay (ELISA) method using commercial kits: leptin, adiponectin, insulin and ghrelin ELISA kits (EMD Millipore Corporation, Billerica, MA, USA, cat. number: #EZRL-83K, #EZRADP-62K, #EZRMI-13K and #EZRGRT-91K, respectively); glucagon EIA kit (Sigma-Aldrich, Saint Louis, MO, USA, cat. number: RAB0202-1KT); and an IGF 1 ELISA kit (Thermo Scientific, Waltham, MA, USA, cat. number: ERIGF1). All serum samples were assayed in duplicate within one assay, and results were expressed in terms of the particular standard hormone. The homeostasis model assessment-β (HOMA-β) was calculated following the formula HOMA-β = (20 × FINS)/(FBG − 3.5); FINS = fasting serum insulin, FGB = fasting blood glucose.