Biotin sp conjugated goat anti rabbit

Histological and immunofluorescence staining on cryosections was performed as described previously (de Melo et al., 2005 (link)). Primary antibodies used were: mouse anti-BrdU (1:200, Chemicon), mouse anti-BRN3A (1:200, Santa Cruz), goat anti-BRN3B (1:200, Santa Cruz), rabbit anti-caspase 3 (1:500, Cell Signaling Technologies), rabbit anti-DLX2 (1:400, C199 affinity purified), mouse anti-ISLET1 (1:600, DSHB, University of Iowa), rabbit anti-phosphohistone H3 (1:1000, Upstate), rabbit anti-PROX1 (1:500, Chemicon), rabbit anti-PAX6 (1:800, Covance) and mouse anti-syntaxin (1:6000, Sigma). Secondary antibodies and fluorescent tertiary molecules used were: FITC-conjugated goat anti-rabbit (1:200), biotin-SP-conjugated goat anti-rabbit (1:200), biotin-SP-conjugated goat anti-mouse (1:200) (Jackson ImmunoResearch), streptavidin-conjugated Oregon Green 488 (1:200) and streptavidin-conjugated Texas Red (1:200) (Molecular Probes). Negative controls omitted the primary antibody. TUNEL staining used the In Situ Cell Death Detection Kit, TMR Red (Roche Diagnostics). Non-radioactive digoxigenin in situ RNA hybridization was performed as described previously (de Melo et al., 2005 (link)).