Rabbit anti rat nrf2 polyclonal antibody

Eight rats in each group were deeply anesthetized at 24 h after injury. Then, brain tissues were rapidly removed and immersed in 4% paraformaldehyde for 72 h. Slices were incubated with 3% H₂O₂ at 37°C for 10 min, normal goat serum for 30 min at room temperature, and primary antibodies of interest at 4°C overnight, and then rinsed three times with 0.1 M phosphate-buffered saline (PBS). Rabbit anti-rat CD34 polyclonal antibody (1:200, Santa Cruz Biotechnology, USA), rabbit anti-rat Nrf2 polyclonal antibody (1:200, Santa Cruz Biotechnology, USA), and goat anti-rat HO-1 polyclonal anti-body (1:200, Santa Cruz Biotechnology, USA) were used to detect protein expression. Then, the slices were incubated with secondary antibodies. Hematoxylin staining was selected as the counterstain. An examiner blinded to the experimental groups detected cell optical density labeled with Nrf2, and HO-1 under a 400× light microscope. The average optical density level of each slice was obtained by Image-Pro Plus 6.0 software (Media Cybernetics, USA)