Rat insulin elisa kit

After 6 weeks, fasted rats (18 h) were anesthetized and sacrificed after 18 h. Serum samples were isolated from 4 ml of venous blood which were drawn from the inferior vena cava and stored at −80 °C for subsequent analysis. Serum glucose, lipids (total cholesterol, HDL cholesterol, triglycerides) and liver enzyme activities of aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP) were determined using the relevant Spinreact kits (Spinreact, Spain). LDL cholesterol was calculated using the Friedwald equation (LDL = total cholesterol – HDL – (triglycerides/5)) [26 (link)]. Serum insulin was determined using Rat-insulin ELISA kit (Merck Millipore,Germany). All serum samples were run in duplicate and analyzed within the same assay. In addition, following sacrifice, the intra-abdominal fat (epididymal, mesenteric and retroperitoneal) was removed from the animals, after which it was cleaned, blotted on a filter paper and weighed.

At the end of the experiment, blood samples were taken, and then animals were sacrificed under anesthesia (Xylasine/Ketamine, 10 mg/90 mg per kg). Kidneys were immediately removed, frozen in liquid nitrogen and stored at −80 °C for later determinations. Plasma insulin level was determined using a rat insulin ELISA kit (EMD Millipore Corporation, USA), calculating the homeostasis model assessment of insulin resistance (HOMA-IR), based on the following formula: $$\text{HOMA}-\text{IR}=\text{serum glucose}\left(\text{mg}/\text{dL}\right)\times \text{plasma insulin}(\mathrm{\mu }\text{U}/\text{mL})/405.$$

Serum insulin was determined using a rat insulin ELISA kit (EMD Millipore Corporation, United States). Total serum testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were measured using a rat ELISA kit (Roche Diagnostics, USA) by applying the manufacturer’s guidelines. The absorbance of colorimetric solutions was assayed using a Utrao microplate reader (Shanghai, China).