Fluorescent confocal image of SY5Y cells stained with Vimentin (S82) antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with Vimentin (S82) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 5 min). Note the highly specific localization of the Vimentin immunosignal to the cytoskeleton, supported by Human Protein Atlas Data (http://www.proteinatlas.org/ENSG00000026025).