Recombinant Anti-c-Fos antibody [EPR21930-238] (ab222699)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21930-238] to c-Fos
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-c-Fos antibody [EPR21930-238]
See all c-Fos primary antibodies -
Description
Rabbit monoclonal [EPR21930-238] to c-Fos -
Host species
Rabbit -
Specificity
In IHC-P, no staining is observed on rat tissues with this antibody in our lab. We recommend ab289723 for work on rat.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Common marmoset -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat and RAW 264.7 grown in serum free medium overnight, followed by treatment with 200 nM PMA for 4 hours, whole cell lysates. IHC-P: Human bladder carcinoma tissue; Mouse dentate gryus and cerebral cortex tissues. ICC/IF: HeLa cells. Flow: HeLa cells. IP: HeLa grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21930-238 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222699 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Detects a band of approximately 55-60 kDa (predicted molecular weight: 41 kDa).
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IHC-P | (4) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/1000.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 55-60 kDa (predicted molecular weight: 41 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
IP
1/30. |
Target
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Function
Nuclear phosphoprotein which forms a tight but non-covalently linked complex with the JUN/AP-1 transcription factor. In the heterodimer, FOS and JUN/AP-1 basic regions each seems to interact with symmetrical DNA half sites. On TGF-beta activation, forms a multimeric SMAD3/SMAD4/JUN/FOS complex at the AP1/SMAD-binding site to regulate TGF-beta-mediated signaling. Has a critical function in regulating the development of cells destined to form and maintain the skeleton. It is thought to have an important role in signal transduction, cell proliferation and differentiation. -
Sequence similarities
Belongs to the bZIP family. Fos subfamily.
Contains 1 bZIP domain. -
Post-translational
modificationsPhosphorylated in the C-terminal upon stimulation by nerve growth factor (NGF) and epidermal growth factor (EGF). Phosphorylated, in vitro, by MAPK and RSK1. Phosphorylation on both Ser-362 and Ser-374 by MAPK1/2 and RSK1/2 leads to protein stabilization with phosphorylation on Ser-374 being the major site for protein stabilization on NGF stimulation. Phosphorylation on Ser-362 and Ser-374 primes further phosphorylations on Thr-325 and Thr-331 through promoting docking of MAPK to the DEF domain. Phosphorylation on Thr-232, induced by HA-RAS, activates the transcriptional activity and antagonizes sumoylation. Phosphorylation on Ser-362 by RSK2 in osteoblasts contributes to osteoblast transformation.
Constitutively sumoylated by SUMO1, SUMO2 and SUMO3. Desumoylated by SENP2. Sumoylation requires heterodimerization with JUN and is enhanced by mitogen stimulation. Sumoylation inhibits the AP-1 transcriptional activity and is, itself, inhibited by Ras-activated phosphorylation on Thr-232. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 2353 Human
- Entrez Gene: 14281 Mouse
- Omim: 164810 Human
- SwissProt: P01100 Human
- SwissProt: P01101 Mouse
- Unigene: 246513 Mouse
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Alternative names
- Activator protein 1 antibody
- AP 1 antibody
- C FOS antibody
see all
Images
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Anti-c-FOS antibody ab222699 was used with Tissue clearing kit – CUBIC (ab316246) and 3D Tissue Staining Kit – CUBIC (ab316248) to penetrate, stain and clear a whole mouse brain. White: nuclear staining, Yellow: c-FOS.
Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain whole brains and get more data from each valuable tissue sample.
For a whole mouse brain, we recommend starting with 0.6 ug of ab222699 and using a Fab fragment secondary antibody at 0.4 ug to create an antibody complex before 3D staining (see protocol for details). Additive A was used during the staining process.
The sample was imaged using a light-sheet microscope.
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Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling c-Fos with ab222699 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in neurons of mouse cerebral cortex (PMID: 24604295). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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c-Fos was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours whole cell lysate with ab222699 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222699 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.
Lane 1: HeLa grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours whole cell lysate 10 μg (Input).
Lane 2: ab222699 IP in HeLa grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222699 in HeLa grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST
Exposure time: 30 seconds.
Serum starvation followed by FBS treatment induces the expression of c-Fos (PMID: 14981092).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permebalized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line grown in serum free medium for 36 hours, followed by addition of 20% FBS for 2 hours (Red) / Untreated control (Green) labeling c-Fos with ab22699 at 1/500 dilution compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution.
Serum starvation followed by FBS treatment induces the expression of c-Fos (PMID: 14981092).
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Immunohistochemical analysis of paraffin-embedded mouse dentate gyrus tissue labeling c-Fos with ab222699 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadic nuclear staining in mouse dentate gyrus (PMID: 24604295). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human bladder carcinoma tissue labeling c-Fos with ab222699 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in tumor cells of human bladder carcinoma (PMID: 28358415). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunofluorescnt analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeablised HeLa (human cervix adenocarcinoma epithelial cell) cells labeling c-Fos with ab222699 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cells in serum free medium for 36 hours, followed by addition of 20% fetal bovine serum for 2 hours (PMID: 14981092).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
Lane 1 : Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) grown in serum free medium overnight, whole cell lysate
Lane 2 : RAW 264.7 grown in serum free medium overnight, followed by treatment with 200 nM PMA for 4 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 41 kDa
Observed band size: 55-60 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
PMA treatment induces expression of c-Fos, as documented in the literature (PMID: 24386331, PMID: 23300800, PMID: 25695333).
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All lanes : Anti-c-Fos antibody [EPR21930-238] (ab222699) at 1/1000 dilution
Lane 1 : Untreated Jurkat (human T cell leukemia cell line from peripheral blood) grown in serum free medium overnight, whole cell lysate
Lane 2 : Jurkat grown in serum free medium overnight, followed by treatment with 200 nM PMA for 4 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 41 kDa
Observed band size: 55-60 kDa why is the actual band size different from the predicted?
Exposure time: 48 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
PMA treatment induces expression of c-Fos, as documented in the literature (PMID: 24386331, PMID: 23300800, PMID: 25695333).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (27)
ab222699 has been referenced in 27 publications.
- Zhang W et al. Reversing the imbalance in bone homeostasis via sustained release of SIRT-1 agonist to promote bone healing under osteoporotic condition. Bioact Mater 19:429-443 (2023). PubMed: 35574058
- Hu B et al. Mesenchymal stem cells-derived exosomal miR-653-5p suppresses laryngeal papilloma progression by inhibiting BZW2. Clinics (Sao Paulo) 78:100129 (2023). PubMed: 36473368
- Chang Y et al. Parvimonas micra activates the Ras/ERK/c-Fos pathway by upregulating miR-218-5p to promote colorectal cancer progression. J Exp Clin Cancer Res 42:13 (2023). PubMed: 36627634
- Zhang Y et al. Endothelin-1, over-expressed in SOD1G93A mice, aggravates injury of NSC34-hSOD1G93A cells through complicated molecular mechanism revealed by quantitative proteomics analysis. Front Cell Neurosci 16:1069617 (2022). PubMed: 36531135
- Wang F et al. Pharmacological mechanisms of Fuzheng Huayu formula for Aristolochic acid I-induced kidney fibrosis through network pharmacology. Front Pharmacol 13:1056865 (2022). PubMed: 36569327