Immunocytochemistry/ Immunofluorescence analysis of neuron/glial cultures labeling Vimentin with ab24525 (green) and GFAP with ab7260 (red). Vimentin is the sole cytoplasmic intermediate filament subunit expressed in fibroblasts, microglial and endothelial cells. The flattened cells in the middle of the image which appear green are fibroblasts. Astrocytes may express primarily GFAP, or both GFAP and vimentin, and so appear red (GFAP only) or golden yellow (GFAP and Vimentin). In cells which express both GFAP and vimentin, the two proteins assemble to produce heteropolymer filaments.

Rat cerebral cortex cultures stained with chicken antibody to vimentin ab24525 (green) and rabbit antibody to GFAP (red). Note flattened fibroblastic cells are mostly green (i.e. vimentin positive, GFAP negative), while clearly astrocytic cells, express both vimentin and GFAP and therefore appear golden or orange. Certain other cells express predominantly GFAP and therefore appear red.

Western blot of Rat whole brain extract,HeLa,SH-SY5Y,HEK293, and NIH/3T3 cells probed with ab24525,showing a single strong band at ~ 50kDa.

ab24525 staining Vimentin in rat smooth muscle cells from mesenteric artery by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 4% paraformaldehyde in physiological saline solution (PSS) 4 min at 4°C and permeabilized with 0.3% Triton x100 before blocking with 2% BSA was done for 30 minutes at 20°C. Samples were incubated with primary antibody (1/300: in PSS with 2%BSA and 0.3% Triton X-100) for 14 hours at 4°C. An Abcam`s ab6875, goat anti-chicken IgY Texas Red was used as secondary antibody at 1/400 dilution.

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