Fluorescent confocal image of SY5Y cells stained with Vimentin antibody. SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated Vimentin primary antibody (1:100, 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).

Western blot of Vimentin antibody in NCI-H460 cell line lysates (35 ug/lane). Vimentin (arrow) was detected using the purified antibody.

Vimentin Antibody flow cytometry of HeLa cells (right histogram) compared to a negative control cell (left histogram). FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.