Rabbit anti tbr1

Immunohistochemical labeling of embryonic brain sections or dissociated neural cells was performed as described previously (Kim et al., 2006 (link)). The following primary antibodies were used: rabbit anti-mTOR (Cell Signaling), rabbit anti-4EBP1 (Cell Signaling), mouse anti-S6 (Cell Signaling), rabbit anti-TSC2 (Cell Signaling), rabbit anti-phospho-mTOR (Cell Signaling), rabbit anti-phospho-4EBP1 (Cell Signaling), rabbit anti-phospho-S6 (Cell Signaling), rabbit anti-phospho-S6K (Cell Signaling), rabbit anti-phospho-histone H3 (Upstate Biotech), mouse anti-BrdU (Sigma), rabbit anti-Ki67 (Covance), rabbit anti-Sox2 (Chemicon), rabbit anti-Tbr1 (Chemicon), rabbit anti-Cux1 (Santa Cruz), goat anti-Brn1 (Novus Biologicals), rabbit anti-Tbr2 (Abcam), mouse anti-Tuj1 (Sigma). Appropriate secondary antibodies conjugated with Alexa Fluor dyes (Invitrogen) were used to detect primary antibodies.

For immunostaining, sections were blocked in 10% Horse Serum, 0.1% Triton-X100 in PBS (PBT) at pH 7.5 for 1 h. Primary antibodies were then diluted in blocking solution and the sections were incubated overnight at 4°C. Primary antibodies included rabbit anti-Tbr1 (1:800, Chemicon; Etobicoke, ON, Canada), rabbit anti-GFP (1:500, Chemicon, Temecula, CA, USA), goat-anti-GFP (1:1000, Abcam) rabbit anti-Pax6 (1:500, Convance), goat anti-Gsx2 (1:500, Millipore), rabbit anti-Tbr2 (1:500, Abcam), rabbit anti-phospho-histone H3 (pHH3; 1:500; Millipore Biotechnology) and rat anti-BrdU (1:20, Serotec). After incubating in primary antibody, the slides were washed three times in PBT and then incubated for 1 h at room temperature in secondary antibodies. Secondary antibodies were conjugated to Alexa568 (1:500; Molecular Probes) or Alexa488 (1:500; Molecular Probes). After incubation with secondary antibodies, the slides were washed three times in PBS and then stained with DAPI (1/10,000 for 5 min) and washed an additional three times. Slides were mounted in Aqua-polymount for imaging.