Immunohistochemical analysis of Birgus latro (Coconut crab) brain tissue, staining Allatostatin 1 (green) with ab53956. The brains were dissected in phosphate buffered saline (0.1 M PBS, pH 7.4) then fixed in 4% PFA in 0.1 PBS, pH 7.4 at 4°C before sectioning at room temperature. The sections were incubated for 90 minutes in PBS-TX (1% BS|A, 0.3% Triton X-100, 0.05% sodium azide, pH 7.4) followed by overnight incubation with primary antibody (1/1000 in PBS-TX) at room temperature. An AlexaFluor®488-conjugated secondary antibody was used to detect staining.