Rabbit anti synaptophysin antibody

Sagittal brain slices, 30 µm thick were used for immunofluorescent staining. The slide containing the tissue sections was fixated in 1:1 acetone isopropanol solution at room temperature for 20 min. The sections were then permeabilized with 0.3% triton-X in PBS containing 0.1% tween 20 (PBS/T). This was followed by a 3X wash with PBS/T. The sections were blocked with 10% v/v normal goat serum (NGS) for 1 h at room temperature. This was followed subsequently by a 3X wash with PBS/T. The sections were incubated overnight at 4 °C with primary antibody diluted to respective concentrations in PBS/T. The primary antibody used were GFAP (GA5) Mouse mAb (1:600; Cell Signaling Tech, 3670 S), Rabbit anti-C1q antibody (1:250; Abcam, ab182451) and Rabbit anti-Synaptophysin antibody (1:600; Abcam, ab52636). On the subsequent day, the sections were incubated with secondary antibodies Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (1:700; Abcam, ab150115) and Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) (1:200; Abcam, ab150089) for 2 h at room temperature. For nuclear staining, ProLong™ Glass Antifade Mountant with NucBlue™ Stain (Sigma Aldrich, product no. 57-50-1) was used. If nuclear staining was not necessary the slides were mounted with DAKO fluorescent mounting media (DAKO, s3023).

In this study, IHC, ICC and Western blots assays utilized the following primary antibodies: Mouse anti-β-actin antibody (Abcam, #ab8226, Cambridge, UK), Mouse anti-GFAP antibody (Cell Signaling Technology, #3670S, Boston, MA, USA), Goat anti-PSD95 antibody (Abcam, #ab12093), Rabbit anti-synaptophysin antibody (Abcam, #ab32127), Rabbit anti-LAMP1 antibody (Abcam, #ab24170), Rabbit anti-MEGF10 antibody (Sigma-Aldrich, #ABC10, St. Louis, MO, USA).