Rabbit monoclonal anti synaptophysin

To examine viral expression pattern at injection and terminal sites, single immunolabeling of neuronal nuclear protein (NeuN) was carried out. Tissue sections were rinsed in 50 mM potassium phosphate buffered saline (KPBS) and incubated in blocking buffer (50 mM KPBS, 0.1% BSA, and 0.2% TritonX-100) for 1 h at room temperature. Sections were then placed in mouse monoclonal anti-NeuN antibody (1:200; Millipore, Billerica, MA) overnight at 4°C. Following incubation, sections were rinsed and placed into Cy5-conjugated goat anti-mouse IgG (1:500; Jackson ImmunoResearch, West Grove, PA) for 30 min. Sections were then rinsed, mounted onto slides, and cover slipped. In order to compare synaptophysin-mCherry expression with synaptophysin immunolabeling, immunohistochemistry for synaptophysin was carried out in a similar manner as above except with rabbit monoclonal anti-synaptophysin (1:200; Abcam, Cambridge, MA) followed by Alexa488-conjugated goat anti-rabbit IgG (1:500; Jackson ImmunoResearch) and DAPI (1 μg/ml for 10 min; Millipore, Billerica, MA).