Milliplex map rat pituitary magnetic bead panel assay kit

Blood samples were collected from the tail in the morning around 10 am and placed at room temperature for about 60 min to coagulate and centrifuged at 1000 g for 10 min. The serum was then removed and stored at −80°C. The samples were analysed using the MILLIPLEX^® MAP rat pituitary magnetic bead panel assay kit (Millipore).

Blood samples were collected from the tail in the morning around 10 am and placed at room temperature for about 60 minutes to coagulate and then centrifuged at 1000 × g for 10 minutes at 5 and 20 weeks after irradiation. The serum was then removed and stored at −80°C. A Luminex assay was used according to the manufacturer's instructions (Millipore, Stockholm, Sweden) to detect pituitary‐related hormones (MILLIPLEX MAP rat pituitary magnetic bead panel assay kit). Reproduction‐related hormone analyses were performed using ultrasensitive rat estradiol, testosterone and progesterone ELISA kits (Crystal Chem, IL, USA). To compare pro‐inflammatory mediator expression profiles between irradiated and non‐irradiated control rats, a Luminex assay was used according to the manufacturer's instructions (Millipore, Stockholm, Sweden). The results were analysed using a Bio‐Plex workstation (Bio‐Rad, Hercules, CA, USA) and normalized to the amount of total protein extracted from hypothalamus homogenates using a colorimetric Bio‐Rad DC Protein Assay kit (Bio‐Rad).