Immunofluorescent confocal-microscopy analysis of mouse cortical section labeling c-Fos with ab208942 at 1/1000 dilution (red) and a rabbit polyclonal anti Fox3/NeuN antibody (green). Neurons positive for c-Fos and Fox3/NeuN appear to be yellow. Inset shows an enlarged image of staining. Nuclei are labeled with Dapi (blue).  

Section of rat hippocampus stained with ab208942 in red and counterstained with rabbit polyclonal antibody to FOX3/NeuN. 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.

Immunohistochemical analysis of mouse brain section (45 μM; fixed by transcardial perfusion with 4% paraformaldehyde) labeling c-Fos with ab208942 at 1/1000 dilution followed by standard HRP-DAB staining technique. Cells expressing c-Fos show dark color in nucleus. Inset shows an enlarged image of ab208942 staining.

All lanes : Anti-c-Fos antibody [2H2] (ab208942) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells in serum free media
Lane 2 : HeLa cells stimulated with 20% fetal bovine serum for 2hrs after 36hrs in serum free media
Lane 3 : Rat cortical neurons
Lane 4 : Rat cortical neurons treated with membrane depolarization buffer for 5hrs

Predicted band size: 41 kDa

Multiple bands at 50-65kDa in stimulated or treated cell lysates  correspond to different forms of the c-Fos proten.

The single band in red at 37 kDa represents GAPDH protein.

All lanes : Anti-c-Fos antibody [2H2] (ab208942) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate (serum-starved for 36 hours)
Lane 2 : HeLa cell lysate (serum-starved and then stimulated with 20% Fetal Bovine Serum for 2 hours)

Predicted band size: 41 kDa

ab208942 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 a monoclonal antibody against GAPDH, used as loading control.

Rat brain neural cultures (left) and the same cells stimulated with membrane deplorization buffer for 5 hours (right).

This is a salt solution containing 170mM Potassium which depolarizes and stimulates gene expression in neuronal cells but has no effect on glia. Cultures were stained with ab208942 in green and rabbit anti-GFAP in red. Nuclear DNA is revealed in blue with the DNA stain DAPI.

Top panel: HeLa (Human epithelial cell line from cervix adenocarcinoma) cells serum starved for 36 hours and then treated with PBS.

Bottom panel: HeLa cells, treated with 20% FBS for 2 hours following serum–starvation for 36 hours, labeling c-Fos using ab208942 at 1/1000 dilution (green).

Red: Vimentin counterstain. Nuclei labeled with DAPI (Blue).