Goat anti ctb antibody

Two hours after the final presentation of the paired stimulus animals were deeply anesthetized with isoflurane and euthanized via transcardial perfusion. After post-fixation and cryoprotection, brains were coronally sectioned (44 um) on a freezing microtome. All tissue for immunohistochemical analysis was processed according to previously described protocols with minor modifications [17 (link),18 (link)]. Representative sections through the rostrocaudal extent of the brain were first mounted and stained for CTb to verify retrograde tracer deposit placement. Free-floating sections were incubated in goat anti-CTb antibody (1:7500; List Biological Laboratories) for 48 hr at 4 °C followed by biotinylated donkey anti-goat IgG (1:1000; Jackson ImmunoResearch Laboratories, West Grove, PA) for 1.5 hr at room temperature, and horseradish peroxidase-conjugated streptavidin (1:1600; Jackson ImmunoResearch Laboratories Inc.) for 1 hr at room temperature. Staining was visualized by addition of 3 % hydrogen peroxide to sections incubating in diaminobenzidine, yielding brown CTb-immunopositive labeling.