Psd 95 antibody

For immunohistochemical analyses, brain sections were rinsed briefly in phosphate-buffered saline and treated with 1% hydrogen peroxide for 15 min. The sections were incubated with a goat anti-Ng antibody (1:500; Abcam) or anti-postsynaptic density protein-95 (PSD-95) antibody (1:500; Abcam). The sections were next incubated with a biotinylated horse anti-goat immunoglobulin G (1:200; Vector Laboratories, Burlingame, CA, USA) and avidin–biotin–peroxidase complex solution and then visualized with a SIGMA FAST^TM 3.3′-diaminobenzidine tablet (Sigma-Aldrich) as a chromogen. To quantify the immunoreactivity of PSD-95, the images were analyzed using the Image-Pro Plus 6.0 program (Media Cybernetics, Rockville, MD, USA). For immunofluorescence staining, brain sections were rinsed briefly in phosphate-buffered saline and incubated with a goat anti-Ng antibody (1:500, 4 °C, 12 h) followed by donkey anti-goat Alexa Fluor^® 594 immunoglobulin G (1:200, room temperature, 1 h; Abcam). All sections were counter-stained with 4′,6-diamidino-2-phenylindole (Thermo Scientific) before mounting.

According to following method (Seo et al., 2019 (link)), the hippocampal tissues were collected, homogenized on ice, and lysed in a lysis buffer. Proteins were quantified using Bradford protein assay (Bio-Rad, Hercules, CA, USA). After electrophoresis, proteins were transferred onto nitrocellulose membrane (GE Healthcare Life Sciences, Chicago, IL, USA). The membrane was blocked with skim milk, then which was treated with mouse β-actin antibody (1:1,000; Santa Cruz Biotechnology), rabbit brain-derived neurotrophic factor (BDNF) antibody (1:1,000; Bioss Antibodies, Woburn, MA, USA), rabbit postsynaptic density 95 kDa (PSD-95) antibody (1:1,000; Abcam, Cambridge, UK), and rabbit synaptophysin (1:1,000; Abcam). Horseradish peroxidase-conjugated anti-mouse for β-actin (1:3,000) and anti-rabbit for BDNF, PDS-95, synaptophysin (1:5,000) were used as secondary antibodies.