Rat leptin elisa kit

Blood was collected from the retro-orbital plexus of overnight fasted rats using microcapillary tubes on 29th day. Serum was separated by centrifugation (4000 rpm for 10 min) and transferred to Eppendorf tubes. The concentrations of lactate dehydrogenase (Reckon Diagnostics Pvt. Ltd., Baroda, India) glucose, total cholesterol, triglycerides (all three from Span diagnostics Ltd., Surat, Gujarat, India), HDL-cholesterol (Reckon Diagnostics Pvt. Ltd., vadodara, India), and apolipoprotein- B by immunoturbidimetric immunoassay kit (Randox Laboratories Ltd., Antrim, UK) in serum were measured with commercial kits. Glycohemoglobin was estimated in blood with a commercial available kit (Asritha In vitro Diagnostic Reagents, Hyderabad, India). The concentrations of insulin and leptin in the serum were measured respectively with rat insulin ELISA kit (Alpco Diagnostics, Salem, USA), rat leptin ELISA kit (BioVendor, Brno, Czech Republic). On the final day of experiment the rats were fasted overnight and sacrificed by cervical dislocation and then different organs (heart, liver, Kidney and pancreas) and fat pads (epididymal, perirenal and mesentric) were removed, washed with normal saline and weighed[12 ].

Biochemical parameters, insulin, leptin and adiponectin were analyzed at the termination of study. The 16-h-fasted rats were etherized by ethyl ether and sacrificed by cardiac exsanguination. Blood samples were placed in centrifuge tube for 30 min and then centrifuged at 3500g for 30 min at 4 °C to obtain serum. After laparotomy, internal organs (liver, kidney and pancreas), epididymal fat and perinephric fat were isolated and weighed. All isolated tissues and serum samples were stored at −80 °C before analysis. Biochemical parameters, such as fasted blood sugar (FBG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, bilirubin, triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) in serum, were measured using a Roche Modular system and all the kits were purchased from Cobas (Roche, Mannheim, Germany). The rat insulin ELISA kit (Mercodia AB), rat leptin ELISA kit (BioVendor) and rat adiponectin ELISA kit (Assaypro) were employed to measure the insulin, leptin and adiponectin, respectively.