Biotin conjugated anti rabbit igg antibody

Protein kinase C gamma (PKCγ) immunohistochemistry was done as previously described (Okada et al., 2019 (link)). Briefly, the brain sections of mice (16–56 weeks old) were incubated twice overnight with anti-PKCγ antibody (1:200; Frontier Institute, Hokkaido, Japan) in 0.5% blocking reagent (Roche Diagnostics, Mannheim, Germany) in PBT at 4°C. The sections were washed with PBT for 15 min six times and then incubated with biotin-conjugated anti-rabbit IgG antibody (1:600; Vector Laboratories, Burlingame, CA, United States) in 0.5% blocking reagent in PBT for 2 h. After being washed with PBT for 15 min five times and with Tris-buffered saline (TBS) supplemented with 0.1% Tween-20 (TBST) for 15 min, the sections were incubated with avidin-biotin peroxidase complex (ABC; Vectastain Elite ABC kit; Vector Laboratories) for 30 min. The sections were washed with 1% Tween-20/TBS for 20 min, and then with TBST for 20 min twice, and were thereafter kept in the same solution at 4°C overnight. The sections were incubated with 3,3′-diaminobenzidine (DAB; Vector Laboratories) for 10 min. All steps were performed at room temperature unless otherwise indicated.