Chicken anti gfp antibody

Immunohistochemistry was used to confirm virus-mediated gene expression in vHipp neurons. Briefly, after behavioral and physiological experiments were performed, rats were transcardially perfused with saline, followed by 4% paraformaldehyde. Brains were post-fixed and cryoprotected in 10% sucrose in phosphate-buffered saline (PBS). For antigen retrieval, free-floating coronal sections from the vHipp (50 μm) were boiled in 10 mM citric acid, pH 6, for a total of 5 min. Sections were then washed in PBS, blocked (2% normal goat serum and 0.3% Triton X-100), then incubated with chicken anti-GFP antibody (Millipore; 1:1000) at 4 °C overnight. Sections were then incubated with AlexaFluor 488 goat anti-chicken secondary antibody (1:1000). To confirm pyramidal cell expression, a subset of sections were then incubated with mouse anti-CAMKII antibody (Thermo Fisher; 1:200) followed by AlexaFluor 594 goat anti-mouse (1:1000). Sections were mounted on slides and cover-slipped using Prolong gold anti-fade reagent. Sections were imaged using an Olympus IX81 Motorized Inverted microscope. The representative images were acquired using FV10-ASW software and enhanced using ImageJ.