Anti-c-Fos 抗体 [2H2]

• IHC-FrFl

Supplier Data

Immunohistochemistry - Free Floating - Anti-c-Fos antibody [2H2] (AB208942)

Section of rat hippocampus stained with ab208942, at a 1/200 dilution, 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [2H2] (AB208942)

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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [2H2] (AB208942)

IHC image of c-Fos staining in a section of formalin-fixed paraffin-embedded normal human placenta* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab208942, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [2H2] (AB208942)

IHC image of c-Fos staining in a section of formalin-fixed paraffin-embedded normal human cervix* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab208942, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [2H2] (AB208942)

IHC image of c-Fos staining in a section of formalin-fixed paraffin-embedded normal human placenta* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was incubated with a Rabbit anti-Mouse IgG1 antibody, ab125913, at 1.5ugml for 15 mins at room temperature and then incubated with ab208942, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [2H2] (AB208942)

IHC image of c-Fos staining in a section of formalin-fixed paraffin-embedded normal mouse brain performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was incubated with a Rabbit anti-Mouse IgG1 antibody, ab125913, at 1.5ugml for 15 mins at room temperature and then incubated with ab208942, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [2H2] (AB208942)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [2H2] (AB208942)

IHC image of c-Fos staining in a section of formalin-fixed paraffin-embedded normal rat placenta performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab208942, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [2H2] (AB208942)

IHC image of c-Fos staining in a section of formalin-fixed paraffin-embedded normal rat brain performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab208942, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Supplier Data

Western blot - Anti-c-Fos antibody [2H2] (AB208942)

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:

Western blot - 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

false

• WB

Supplier Data

Western blot - Anti-c-Fos antibody [2H2] (AB208942)

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.

All lanes:

Western blot - 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

false

• WB

Lab

Western blot - Anti-c-Fos antibody [2H2] (AB208942)

False colour image of Western blot : Anti-c-Fos antibody [2H2] staining at 1/1000 dilution, shown in green; Rabbit anti-ACTN4 [EPR2533(2)] (ab108198) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab208942 was shown to bind specifically to c-Fos. A band was observed at 45-55 kDa in treated wild-type PC-12 cell lysates with no signal observed at this size in FOS knockout cell line ab281615 (knockout cell lysate ab283111). Treatment of NGF has no observable affect on protein expression in this cell line. To generate this image, wild-type and FOS knockout PC-12 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-c-Fos antibody [2H2] (ab208942) at 1/1000 dilution

Lane 1:

Wild-type PC-12 Untreated Control NGF (0 ng/mL, 4 days) cell lysate at 20 µg

Lane 2:

Wild-type PC-12 Treated NGF (111 ng/mL, 4 days) cell lysate at 20 µg

Lane 3:

FOS knockout PC-12 Untreated Control NGF (0 ng/mL, 4 days) cell lysate at 20 µg

Lane 4:

FOS knockout PC-12 Treated NGF (111 ng/mL, 4 days) cell lysate at 20 µg

Observed band size: 45-55 kDa

false