Mouse monoclonal anti glial fibrillary acidic protein antibody

The tissue sections were subjected to microwave heat-induced epitope retrieval. After blocking with 4% goat serum in PBS containing 0.3% Triton X-100 for 1 hr, the sections were incubated with primary antibodies at 4℃ overnight. The primary antibodies included rat polyclonal anti-BrdU antibody (1:1,000; Abcam, UK), rabbit anti-doublecortin (DCX) polyclonal antibody (1:1,000; Abcam, UK), mouse monoclonal anti-neuron-specific nuclear protein (NeuN) antibody (1:1,000; Chemicon, USA), and mouse monoclonal anti-glial fibrillary acidic protein (GFAP) antibody (1:800; Sigma, St. Louis, MO, USA). The sections were then washed with PBS and incubated with secondary antibodies [fluorescein isothiocyanate (FITC) or tetramethylrhodamine isothiocyanate (TRITC)-conjugated IgG, or biotinylated IgG] for 2 hr at room temperature. To detect apoptosis of newly generated cells in the SGZ, the BrdU-stained sections were labeled using the terminal dUTP nick-end labeling (TUNEL) assay, according to the instruction of in situ Cell Death Detection Kit (Roche, Germany), and negative/positive controls were also used according to the instructions. A laser confocal microscope (TSC SP2, Leica, Manheim, Germany) detected the fluorescence signals.