Fluorescent confocal image of SH-SY5Y cells stained with Nestin (S1409) antibody. SH-SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0. 2%, 30 min). Cells were then incubated with A00806-1 Nestin (S1409) 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 μg/ml, 5 min). Note the highly specific localization of the Nestin (S1409) immunosignal to the intermediate filaments, supported by Human Protein Atlas Data (http://www. proteinatlas. org/ENSG00000132688).