Bace1 antibody

For BACE1 immunohistochemistry, cross sections of the retina and brain were incubated with 50% formamide and 50% sodium citrate buffer for 1 hour in a 65°C water bath for antigen retrieval. Retinal sections were soaked in 1% H₂O₂ for 30 minutes at room temperature to abolish endogenous peroxidase activity. After rinsing with PBS, sections were treated with a blocking solution containing 5% bovine serum albumin and 0.1% Triton X-100 for 1 hour at room temperature. Then, sections were stained overnight at 4°C with a rabbit polyclonal BACE1 antibody (1:100; Abcam, Hong Kong, China) diluted in blocking solution. Thereafter, sections were rinsed in PBS/0.1% Triton X-100 for 5 minutes, and then incubated with goat anti-rabbit secondary antibody (1:200; Vector, Burlingame, CA, USA) for 2 hours at room temperature. Sections were washed with PBS/0.1% Triton X-100 for 5 minutes, and then incubated with avidin-biotin-peroxidase complex (1:200; ABC Standard Elite, Vector Laboratories, Burlingame, CA, USA) for 1 hour at room temperature. Staining was evaluated using conventional optical microscopy (Olympus, Tokyo, Japan) after the DAB color reaction. In control sections, the primary antibody was omitted.