Somatostatin

Brain sections were immunostained for somatostatin (Santa Cruz Biotechnology, Santa Cruz, CA; catalog number SC7819) as described in Spiegel et al. (2013) (link). Briefly, free-floating sections were washed in 0.1 M phosphate-buffered saline (PBS), and then endogenous peroxidases were quenched in 0.3% hydrogen peroxide in PBS. After additional PBS washes, sections were blocked in 5% normal horse serum in PBS with 0.3% Triton. Sections were then incubated with primary antibody at a dilution of 1:1,600 in PBS containing 0.15% Triton and 3% normal serum for 72 hr at 4°C with agitation. Following primary antibody incubation, sections were washed in PBS and reacted with biotinylated secondary antibody horse antigoat IgG (Vector Laboratories, Burlingame, CA) diluted in PBS with 0.15% Triton and 5% normal horse serum for 45 min. The secondary antibody was detected with avidin-biotin complex (ABC Elite; Vector Laboratories) and the avidin-biotin complex was visualized with nickel-enhanced diaminobenzadine (Vector Laboratories). Tissue sections were mounted onto gelatin coated slides and dried, dehydrated with increasing ethanol concentrations, cleared in xylene, and coverslipped using DPX mounting media.