Rabbit sv2a antibody

Total hippocampal proteins were extracted, separated by polyacrylamide gel electrophoresis, and then transferred to polyvinylidene fluoride membranes. The membranes were blocked with 5% nonfat milk for 2 h at room temperature, washed three times with tris‐buffered saline Tween (TBS‐T) solution, mixed with primary antibody (rabbit SV2A antibody, Abcam, UK; 1:1000 dilution), and incubated overnight at 4°C. Then, the membrane was washed three times with TBS‐T and incubated with horseradish peroxidase‐conjugated secondary antibody (goat anti‐rabbit IgG antibody, Bioss, Shanghai, China; 1:10000 dilution) at room temperature for 90 min. The membrane was washed thrice with TBS‐T solution and the protein bands were detected using enhanced chemiluminescence (ECL) kit (Beyotime Biotechnology, Shanghai, China). Finally, the protein band intensities were quantified using ImageJ software.

Frozen sectioning was used to slice brain samples from AD patients and non-AD patients; samples were donated by the Department of Anatomy, Histology, and Embryology (School of Basic Medicine, Fudan University, Shanghai, China). Sections (20 μm) were rinsed three times with PBS and then permeabilized with 0.1% Triton X-100 in PBS, blocked with 5% BSA in PBS at RT for 30 min, and incubated overnight at RT with both rabbit SV2A antibody (1 μg/ml, Abcam) and mouse APP antibody (1 μg/ml, Covance). Subsequently, sections were washed three times in PBS, followed by a 2-h incubation with goat anti-rabbit IgG (594; 3 μg/ml, Abcam) and goat anti-mouse IgG (488; 3 μg/ml, Abcam). Sections were again washed three times with PBS. Fluorescence intensity was detected by using a Zeiss LSM710 fluorescence microscope (Liu et al., 2018 (link)).