Rat insulin enzyme linked immunosorbent assay elisa kit

Fasting blood glucose (FBG) and postprandial blood glucose (PBG) levels were measured each week during the experimental period using a blood glucometer (One Touch^®UltraVue, Johnson & Johnson Medical Ltd., Shanghai). Serum glycosylated hemoglobin A1c (HbA1c) levels were estimated by the method described by Wolfram S, et al.,^{10 (link)} and muscle and hepatic HbA1c levels were determined according to the method provided by Macauley et al.¹¹ Fasting serum insulin (FSI) levels were evaluated using a rat insulin enzyme-linked immunosorbent assay (ELISA) kit (Thermo Fisher Scientific, Waltham, USA) according to the manufacturer’s protocol. Homeostasis model of insulin resistance (HOMA-IR) is the gold standard for estimating insulin resistance and is derived using the following equation¹¹ $HOMA-IR=FBG\left(mmol/L\right)\times FSI\left(mU/L\right)]/22.5.$

A subset of the blood of collected, CON (n = 9) and CAF (n = 9) were used for serum analyses. Glucose, triglycerides (TAGs), total cholesterol (T-CHOL) and high-density lipoprotein (HDL-) cholesterol levels were analysed using colorimetric kits according to manufacturer's instructions (Human Diagnostics, Wiesbaden, Germany). LDL- and VLDL-cholesterol were calculated according to Friedewald, Levy, and Fredrickson [14] (link). Insulin was assessed using a Rat Insulin Enzyme Linked Immunosorbent Assay (ELISA) kit (Thermo Scientific, MD, USA). Inflammatory status – cytokines and/or chemokines, namely interleukin (IL) 1α and β, IL2, IL4, IL6, IL10, IL12, IL13, interferon (IFN)-γ, tumour necrosis factor (TNF)-α, granulocyte-macrophage colony-stimulating factor (GM-CSF), and regulated on activation, normal T cell expressed and secreted (RANTES) – was profiled using a Rat Inflammatory Cytokines Multi-Analyte ELISArray kit (Qiagen, MD, USA) for the week 18 blood samples only. The absorbance was read and the fold changes were assessed by determining the ratio between the absorbance values of the two groups of each cytokine/chemokine.