Rabbit anti vglut1

Mouse anti-β-Tubulin (1:2000, Cell Signaling Technology), rabbit anti-GAPDH (1:2000, Cell Signaling Technology), mouse anti-Calretinin (1:500, Swant), mouse anti-Calbindin (1:500, Swant) rabbit anti-Myelin Basic Protein (1:1000, Abcam), rabbit anti-VGLUT1 (1:1000, Millipore). Secondary antibodies used for this study were: HRP conjugated donkey anti-mouse (1:2000, Jackson ImmunoResearch) and donkey anti-rabbit (1:2000, Jackson ImmunoResearch).

Immunofluorescence on coverslip cultures was performed according to previous reports (Yan et al., 2005 (link)). Cells were fixed in 4% paraformaldehyde in PBS for 20 min and blocked in blocking solution (PBS containing 10% normal donkey serum and 0.2% Triton X-100) for 1 h. The following primary antibodies were used: mouse anti-SSEA-3 (1:200), mouse anti-SSEA-4 (1:400), mouse anti-Nkx6.1 (1:100), mouse anti-En1 (1:100) (DSHB, Iowa City, USA), anti-TRA-1-60 (1:150), anti-TRA-1-81 (1:150), mouse anti-Synaptophysin (1:1000), rabbit anti-v-Glut1 (1:2000) (Millipore, Billerica, USA), anti-Nanog (1:150), goat anti-Otx2 (1:500) (R&D), mouse anti-Tuj1 (1:5000) (Abcam, Cambridge, UK), rabbit anti-MAP2 (1:1000), mouse anti-TH (1:1000) (Sigma), mouse anti-HB9 (1:50) and goat anti-FoxA2 (1:500) (Santa Cruz, California, USA). After an overnight incubation at 4°C, species-specific secondary antibodies conjugated with Alexa Fluor 488 or 594 or CY5 (1:1000) (Invitrogen) were applied for 1 h at room temperature. Cell nuclei were stained with DAPI (Sigma). Images were obtained using a Leica TCS SP8 confocal laser-scanning microscope (Leica, Wetzlar, Germany).