Anti-c-Fos antibody [N486/32]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [N486/32] (AB302667)

Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling c-Fos with ab302667 at 1/500 (1.676 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human cervical cancer.The section was incubated with ab302667 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody (610-4141) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [N486/32] (AB302667)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling c-Fos with ab302667 at 1/500 (1.676 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human colon.The section was incubated with ab302667 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody (610-4141) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [N486/32] (AB302667)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling c-Fos with ab302667 at 1/250 (3.4 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing increased nuclear staining in HeLa cells after starvation 16 hours then treated with TPA (200 nM) for 4 h is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [N486/32] (AB302667)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling c-Fos with ab302667 at 1/250 (3.4 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing increased nuclear staining in NIH/3T3 cells after starvation 16 hours then treated with TPA (200 nM) and MG-132(10 µM) for 4 h is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [N486/32] (AB302667)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling c-Fos with ab302667 at 1/2000 (0.419 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on mouse cerebrum.The section was incubated with ab302667 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody (610-4141) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Fos antibody [N486/32] (AB302667)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling c-Fos with ab302667 at 1/1000 (0.838 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on rat cerebrum.The section was incubated with ab302667 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody (610-4141) for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-c-Fos antibody [N486/32] (AB302667)

Data of BM-Recombinant Anti-c-Fos antibody [EPR20769] (ab214672) was in the internal.Blocking and diluting buffer and concentration : 5% NFDM/TBST The band(s) beneath the target band are likely to be degraded target fragments (PMID : 9737957 and PMID : 20498278). Exposure time : Lanes 1-2 : 26 seconds Lanes 3-4 : 81 seconds Lanes 5-6 : 136 seconds

All lanes:

Western blot - Anti-c-Fos antibody [N486/32] (ab302667) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) starved overnight, whole cell lysate 20 µg

Lane 2:

HeLa starved overnight, then treated with 200nM PMA (<a href='/ko/products/biochemicals/phorbol-12-myristate-13-acetate-pma-pkc-activator-ab120297'>ab120297</a>) for 4 hours, whole cell lysate 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) starved overnight, whole cell lysate 20 µg

Lane 4:

NIH/3T3 starved overnight, then treated with 200nM PMA (<a href='/ko/products/biochemicals/phorbol-12-myristate-13-acetate-pma-pkc-activator-ab120297'>ab120297</a>) and 10uM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 4 hours, whole cell lysate for 4 hours, whole cell lysate 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma) starved overnight, whole cell lysate 20 µg

Lane 6:

PC-12 starved overnight, then treated with /ml NGF (<a href='/ko/products/proteins-peptides/recombinant-human-ngf-protein-active-ab9796'>ab9796</a>) and 10uM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 2 hours, whole cell lysate 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 37 kDa,55 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-c-Fos antibody [N486/32] (AB302667)

Western blot : Anti-c-Fos antibody [N486/32] ab302667 staining at 1/1000 dilution, shown in green; Rabbit anti-ACTN4 ab108198 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 35-50 kDa in Wild-type A375 cell lysates with no signal observed at this size in FOS knockout A375 cell line. To generate this image, 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 800CW and Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-c-Fos antibody [N486/32] (ab302667) at 1/1000 dilution

Lane 1:

Wild-type A375 cell lysates at 20 µg

Lane 2:

FOS knockout A375 cell lysates at 20 µg

Lane 3:

Wild-type PC-12 ab279978 cell lysates at 20 µg

Lane 4:

Western blot - Rat FOS (c-Fos) knockout PC-12 cell line (ab281615) at 20 µg

Lane 5:

HeLa cell lysates at 20 µg

Lane 6:

Ramos cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 35-50 kDa

false