Human adiponectin elisa

The concentrations of the adipokines, adiponectin and leptin in serum samples were assayed at the “Nutrition Research Unit” Laboratory at King Fahd Medical Research Center. Serum adiponectin was measured using “Biovendor human adiponectin ELISA” high sensitivity kit, while serum leptin was assayed using DRG®Leptin ELISA kit. Both assays were solid phase enzyme-linked immunosorbent assay, and were carried out according to the manufacturers’ procedure. Absorbance was measured at 450±10 nm using a microplate reader (Biokit, ELX800- USA).
Serum biochemical and remaining endocrine parameters were assayed in accredited Clinical Chemistry laboratory at the National Guard Hospital, King Abdulaziz Medical City-Jeddah. For serum glucose, liver enzymes, albumin and total protein tests Abbott architect c16000 auto-analyzer (spectrophotometric method) was used. On the same auto-analyzer, the method of immunoturbidimetric determination technique was utilized to measure hsCRP. Insulin measurement was done on Abbott Architect i2000 auto-analyser by using the method of a chemiluminescent microparticle immunoassay (CMIA). Growth hormone was measured by using chemiluminescent immunometric method on immulite 1000 analyzer.

All patients were treated according to international recommendations for hyperglycemia, with appropriate amounts of intravenous fluid and intravenous crystalline insulin infusion. Euglycemia was reached within 6–12 h. Serum samples were taken from the patients on admission and at 6 h (which was chosen as an optimum time for half-life of adiponectin) after normalization of blood glucose to measure adiponectin levels. Adiponectin levels were measured by Biovendor Human Adiponectin ELISA. Results are given as μg/mL.