Rabbit antibody to gfp

The generation, dissection, immunostaining and mounting of mosaic clones in adult brains have been described^{12 (link),33 (link)}. For MARCM experiments, mosaic clones were induced from NHL to 96 h ALH by heat-shock for 15–40 min. For twin-spot MARCM experiments, mosaic clones of embryonic-born adPNs were generated by heat-shock for 12 min. Primary antibodies used in this study included rat monoclonal antibody to mCD8 (1:100, Invitrogen), chicken antibody to GFP (1:800, Invitrogen), rabbit antibody to GFP (1:800, Invitrogen), rabbit antibody to RFP (1:800, Clontech), rabbit antibody to γ-aminobutyric acid (GABA; 1:100, Sigma), rabbit antibody to phospho-Histone H3 (H3-P; 1:200, Millipore), nc82 (1:100, Developmental studies hybridoma bank/DSHB) and mouse antibody to choline acetyltransferase (Chat; 1:100, DSHB). Secondary antibodies with different fluorophores, including Alexa 488, 546 and 647 (Invitrogen), were used at 1:800 dilutions in this study. Immunofluoroscence images were collected by Zeiss LSM 700 or 780 confocal microscopy and further processed using Adobe Photoshop. The plugin “Cell Counter” and the analyzing tool “ROI Manager” from Fiji ImageJ were used to count neuronal number and estimate the SIP1::sfGFP expression, respectively. One-way ANOVA with post-hoc Tukey test was used for statistical analysis in this study.

Primary antibodies and dilutions used were: rat antibody to CTIP2 (1:500, Abcam); mouse antibody to SATB2 (1:200, Abcam); rabbit antibody to SIRT1 (1:250, Millipore); rabbit antibody to CTIP1 (1:500, Abcam); rabbit antibody to GFP (1:500, Invitrogen); rabbit antibody to Ki67 (1:500, Abcam); chicken antibody to NESTIN (1:500, Novus Biologicals); mouse antibody to TuJ1 (1:500, Covance); and mouse antibody to MAP2 (1:500, Sigma). Alexa fluorophore conjugated secondary antibodies from Invitrogen were used at a dilution of 1:1000. Hoechst 33342 counterstain was used to visualize nuclei (1:3,000, Invitrogen).