Insulin ria kit

The concentrations of blood serum hormones involved in the regulation of feed intake and catabolic/anabolic pathways and carbohydrate/lipid/protein turnover were investigated in the blood serum as follows using commercial RIA and EIA kits: insulin (Insulin RIA kit, Millipore, USA), total thyroxine (Thyroxine (T4) kit, RIA – Cis International), free thyroxine (Free Thyroxine (fT4) kit, RIA – Cis International), total triiodothyronine (Triiodothyronine (T3) kit, RIA – Cis International), free triiodothyronine (Free Triiodothyronine (fT3) kit, RIA – Cis International), corticosterone (Corticosterone ELISA kit – Enzo Life Sci., Warsaw, Poland), leptin (Multi-Species Leptin RIA, Millipore, USA), incretins GIP (Gastric Inhibitory Polypeptide) (SunRed, Jufengyuan Road, Baoshan, District, Shanghai), and GLP-1 (Glucagon-like Polypeptide-1) (Phoenix Pharmaceuticals Inc., USA). In order to verify the specificity of binding of kits, serum dilution curve was performed. For this purpose serum from five randomly chickens was diluted 2×, 4× and 10 times in Elisa/RIA buffer, and next concentrations of investigated hormones were determined in all dilutions.

The blood serum hormone concentration was analyzed using commercial RIA and EIA kits: insulin (Insulin RIA kit, Millipore, Billerica, Massachusetts, USA), leptin (Multi-Species Leptin RIA, Millipore), glucagon (EIA kit, Phoenix Pharmaceuticals Inc., Burlingame, CA, USA) as described by Kolodziejski et al. [24 (link)]. To verify the binding specificity of the kits, a serum dilution curve was performed. For this purpose, serum from five randomly selected individuals per group was diluted 2 times, 4 times, and 10 times in ELISA/RIA buffer, and the concentrations of investigated hormones were determined in all dilutions. Glucose was measured using a Glucose Assay Kit (Pointe Scientific, Canton, Michigan, USA).

Terminal blood collection after an overnight fast was performed following skeletal muscle extraction via cardiac puncture. A glucometer reading (OneTouch Ultra 2, LifeScan Inc., Milpitas, CA, USA) of whole blood glucose was made at this time. Blood samples were collected in heparinized tubes and centrifuged at 9 300 g for 5 minutes at 4°C. Plasma was stored at −80°C for analysis of glucose (via glucometer) and insulin (rat insulin RIA kit; Millipore, St. Charles, MO, USA).