Anti-c-Fos Antibody [2H2] (A85387)

Section of rat hippocampus stained with Anti-c-Fos Antibody (red) and Anti-FOX3/NeuN Antibody (A85403 | green). DAPI reveals nuclei of neurons and glia in blue. The hippocampal neurons stain green for FOX3/NeuN and a few also are expressing c-FOS, and so appear orange. These cells were spontaneously active at the time the animal was sacrificed.

Left: Anti-c-Fos Antibody staining (green) in HeLa cells, which were treated with serum-starvation for 36 hours, followed by 2 hours, 20% FBS stimulation (bottom panel), or PBS treatment (top panel). Green c-Fos staining only localizes in the nuclei of stimulated cells, but not in un-stimulated cells. Cells are counter-stained with Anti-Vimentin Antibody (A85421 | red). Blue shows DAPI staining of nucleus. Middle: Mouse brain section (45 µM; fixed by transcardial perfusion with 4% paraformaldehyde) labeled with Anti-c-Fos Antibody using a standard HRP-DAB (horseradish peroxidase-3,3’-diaminobenzidine) staining technique. Cells expressing c-Fos show dark color in nucleus. Right: Mouse cortical section labeled with Anti-c-Fos Antibody (red) and Anti-Fox3/NeuN Antibody (A85403 | green) using immuno-fluorescent confocal-microscopy. Neurons positive for c-Fos and Fox3/NeuN appear to be yellow. Inset shows an enlarged image of Anti-c-Fos Antibody staining. Nuclei are labeled with DAPI (blue).

Western blot analysis of cell lysates using Anti-c-Fos Antibody [2H2] (A85387), at a dilution of 1:1,000, in green, and Anti-GAPDH Antibody (A85377), at a dilution of 1:20,000, in red, used as a loading control. The lanes contain samples of: [1] Protein standards, in red, [2] HeLa cells in serum free media, [3] HeLa cells stimulated with 20% fetal bovine serum for 2hrs after 36hrs in serum free media, [4] rat cortical neurons, and [5] rat cortical neurons treated with membrane depolarization buffer for 5hrs. Multiple bands at 50-65kDa in stimulated or treated cell lysates correspond to different forms of the c-Fos proten. The single band at 37 kDa represents GAPDH protein.

Top panel: Western blot analysis of c-Fos expression in HeLa cells using Anti-c-Fos Antibody. Lane 1: HeLa cells were serum-starved for 36 hours. 2: Serum-starved HeLa cells were stimulated with 20% FBS (fetal bovine serum) for 2 hours. Anti-c-Fos Antibody recognizes bands in the range of 50-65 kDa, which represent multiple forms of c-Fos. Serum starvation attenuates c-Fos expression, while 20% FBS strongly stimulates c-Fos expression. Bottom panel: Blot was stripped and probed with Anti-GAPDH Antibody (A85382), used as loading control.