Immunohistochemical analysis of mouse RPE/Choroidal whole-mounts labeling VEGF with ab14078 at 1:100 dilution. Left panel: purple staining VEGF. Right panel: overlay with Iba1 antibody. VEGF expression is shown in accumulating macrophages following laser induced choroidal neovascularisation.    Eyes were enucleated and fixed in 2% (wt/vol) paraformaldehyde (PFA). After dissection of RPE/choroid, tissues were blocked and permeabilized in 5% BSA, with 0.3% Triton X-100 in PBS for 2 hours, followed by incubation with antibodies in 1% BSA with 0.15% Triton X-100 at 4°C overnight. After thorough washing, samples were further incubated with secondary antibodies at RT for 3 hours in the dark. Tissues were washed and flat-mounted in medium.