Rabbit anti catalase cat

In brief, according to our published method (19 (link)), sheep anti-superoxide dismutase 1 (SOD1) (1:1,000; EMD Millipore, Billerica, MA, USA), sheep anti-mitochondrial (SOD2) (1:1,000; EMD Millipore), rabbit anti-catalase (CAT) (1:500; EMD Millipore), mouse anti-glutathione peroxidase (GPX) (1:500; EMD Millipore), mouse anti-NeuN (a marker for neuron) (1:1,000; Thermo Fisher Scientific, Inc., Waltham, MA, USA), mouse anti-GFAP (a marker for astrocyte) (1:800; Abcam, Cambridge, MA, USA), and rabbit anti-Iba1 (a marker for microglia) (1:800; Wako Pure Chemical Industries, Ltd., Osaka, Japan) were used as primary antibodies. The sections were sequentially treated with 0.3% hydrogen peroxide (H₂O₂) in PBS for 30 min and 10% normal goat serum in 0.05 M PBS for 30 min. The treated sections were incubated with the primary antibodies overnight at 4°C, thereafter, the reacted sections were exposed to biotinylated goat anti-mouse or goat anti-rabbit IgG (1:200; Vector Laboratories, Inc., Burlingame, CA, US) and streptavidin peroxidase complex (1:200; Vector Laboratories, Inc.). Finally, the reacted sections were visualized by staining with 3, 3′-diaminobenzidine tetrahydrochloride in 0.1 M Tris-HCl buffer (pH 7.2).

Immunohistochemistry was performed according to the previously published procedure (24 (link)). In short, the sections were incubated with diluted mouse anti-neuronal nuclei (NeuN; 1:800; cat. no. MAB377), sheep anti-copper, zinc-superoxide dismutase (SOD1; 1:1,000; cat. no. 574597) and sheep anti-mangan-superoxide dismutase (SOD2; 1:1,000; cat. no. 574596; all from EMD Millipore, Billerica, MA, USA), rabbit anti-catalase (CAT; 1:500; cat. no. ab52477) and sheep anti-glutathione peroxidase (GPX; 1:1,000; cat. no. ab21966; both from Abcam, Cambridge, MA, UK). Thereafter the tissues were exposed to biotinylated horse anti-mouse (1:250; cat. no. BA-2000), rabbit anti-sheep (1:250; cat. no. BA-6000) and goat anti-rabbit immunoglobulin (Ig)G (1:250; cat. no. BA-1000) and streptavidin peroxidase complex (1:200, all from Vector Laboratories, Burlingame, CA, USA) and were visualized with 3,3′-diaminobenzidine (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).