Lh elisa kit

Blood samples were collected and allowed to clot overnight at 4°C before being centrifuged for 20 minutes at 1000g. The supernatant was subsequently collected, and enzyme immunoassays (EIAs) for GnRH (S-1217, LHRH-EIA Kit, Peninsula Laboratories International Inc., San Carlos, CA, USA), LH (LH ELISA Kit, ENZ-KIT107, Enzo Life Sciences Inc., Berne, Switzerland), FSH (Rat FSH ELISA Kit, abx051243, Abbexa, Cambridge, UK), and testosterone (T) (Testosterone ELISA Kit, ADI-900-065, Enzo Life Sciences Inc.) were conducted.

Blood samples were collected directly from the inferior vena cava with a 1-mL syringe at the end of the experiment. Serum was obtained by centrifugation at 13,000 rpm for 15 min and stored at −70 °C until use. Serum luteinizing hormone (LH) levels were measured using LH ELISA kit (Enzo Life Science, Inc., Farmingdale, NY, USA). Serum follicle-stimulating hormone (FSH) levels were measured using an FSH ELISA kit (Enzo Life Science, Inc., Farmingdale, NY, USA). Serum 17β-Estradiol levels were measured using a 17β-Estradiol ELISA kit (Enzo Life Science, Inc., Farmingdale, NY, USA). Serum testosterone levels were measured using a testosterone ELISA kit (Enzo Life Science, Inc., Farmingdale, NY, USA). All kits were used according to the manufacturer’s instructions, and each sample was analyzed in duplicate.

Throughout the entire experimental period, weekly blood samples were obtained from tail vein of all rats. Serum was obtained and kept at −20°C until hormonal measurements. The concentrations of estradiol (E2) (17β-Estradiol high sensitivity ELISA kit; Catalog #: ADI-900–174) and luteinizing hormone (LH) (LH ELISA KIT; Catalog #: ENZ-KIT107) were measured using ELISA kits (Enzo Life Science) following the protocols provided by the manufacturer.