Mouse anti tuj1 antibody

Immunostaining was performed as previously described [26 (link)]. Briefly, NSCs or E16.5 mouse cortical neurons were seeded onto an 8-well chamber. The neurons were fixed with 4% paraformaldehyde in phosphate buffered saline (PBS) for 15min and permeabilized with 0.2% TritonX-100 in PBS for 10min. After being blocked with 5% Normal Goat Serum (Thermo Fisher) at room temperature (RT) for 1h, the cells were incubated with mouse anti-Nestin antibody (Millipore, MAB353) and rabbit anti-Sox2 antibody (Abcam, ab97959) for NSCs, or with mouse anti-Tuj1 antibody (Biolegend, 801,201) and rabbit anti-GFAP antibody (Sigma, HPA056030) for E16.5 cortical neurons at 4°C overnight. Then, the cells were incubated with Cy3 conjugated anti-rabbit IgG (A10520, Invitrogen) and Alexa Fluor 488 conjugated anti-mouse IgG (A10680, Invitrogen) secondary antibodies at RT in darkness for 1h. After washing 3 × 5min with 1 × PBS, cells were mounted with DAPI-Fluoromount-G^™ Clear Mounting Media (Southern Biotech, 010020). Fluorescent images were acquired using a confocal microscope.