Rabbit anti 5 ht antibody

Immunohistochemistry for 5-HT and TPH was performed according to the previously described method (Kim et al., 2015 (link); Roh et al., 2016 (link)). Briefly, the sections were incubated in PBS for 10 min and washed 3 times, again with PBS and then incubated in 1% H₂O₂ for 30 min. Next, the sections were incubated overnight with rabbit anti-5-HT antibody (Immunostar, Hudson, NY, USA) at a dilution of 1:500 for visualization of 5-HT expression or with mouse anti-TPH antibody (Oncogene Research Product, Cambridge, UK) at a dilution of 1:1,000 for visualization of TPH expression. The sections were then incubated for 1 hr with biotinylated anti-rabbit secondary antibody or with anti-mouse secondary antibody (Vector Laboratories, Burlingame, CA, USA). The sections were subsequently incubated with avidin-biotin-peroxidase complex (Vector Laboratories) for 1 hr at room temperature. Immunoreactivity was visualized by incubating the sections in a solution consisting of 0.05% 3,3′-diaminobenzidine and 0.01% H₂O₂ in 50-mM Tris-buffer (pH, 7.6) for approximately 3 min. The sections were then mounted on gelatin-coated glass slides. The slides were air-dried overnight at room temperature and dehydrated through a gradient of ethanol and covered with coverslips using Permount (Fisher Scientific Inc.).

Immunohistochemistry for 5-HT and TPH was performed according to the previously described method (Seo et al., 2011 (link); Sung et al., 2010 (link)). Briefly, the sections were incubated in PBS for 10 min and washed 3 times, again with PBS and then incubated in 1% H₂O₂ for 30 min. Next, the sections were incubated overnight with rabbit anti-5-HT antibody (Immunostar, Hudson, NY, USA) at a dilution of 1:500 for visualization of 5-HT expression or with mouse anti-TPH antibody (Oncogene Research Product, Cambridge, UK) at a dilution of 1:1,000 for visualization of TPH expression. The sections were then incubated for 1 hr with biotinylated antirabbit secondary antibody or with antimouse secondary antibody (Vector Laboratories). The sections were subsequently incubated with avidin-biotin-peroxidase complex (Vector Laboratories) for 1 hr at room temperature. Immunoreactivity was visualized by incubating the sections in a solution consisting of 0.05% DAB (Sigma Chemical Co.) and 0.01% H₂O₂ in 50-mM Tris-buffer (pH, 7.6) for approximately 3 min. The sections were then mounted on gelatin-coated glass slides. The slides were air-dried overnight at room temperature and dehydrated through a gradient of ethanol and covered with coverslips using Permount (Fisher Scientific).