Recombinant
RabMAb

Recombinant Anti-c-Fos antibody [EPR20769] (ab214672)

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Overview

  • Product name

    Anti-c-Fos antibody [EPR20769]
    See all c-Fos primary antibodies

  • Description

    Rabbit monoclonal [EPR20769] to c-Fos

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    This product was produced with the following immunogens:
    Recombinant fragment within Human c-Fos aa 1-100. The exact sequence is proprietary.
    Database link: P01100

    Recombinant fragment within Human c-Fos aa 200-300. The exact sequence is proprietary.
    Database link: P01100

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • WB: RAW264.7, Jurkat (PMA treated) and HeLa (20% FBS 2 hours) whole cell lysates. ICC/IF: HeLa cells.

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214672 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
IP 1/40.
ICC/IF 1/500.
WB 1/1000. Detects a band of approximately 55-60 kDa (predicted molecular weight: 40 kDa).

Target

  • 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 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
    modifications

    Phosphorylated 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

  • Database links

    • Alternative names

      • Activator protein 1 antibody
      • AP 1 antibody
      • C FOS antibody
      • Cellular oncogene c fos antibody
      • Cellular oncogene fos antibody
      • FBJ murine osteosarcoma viral (v fos) oncogene homolog (oncogene FOS) antibody
      • FBJ murine osteosarcoma viral oncogene homolog antibody
      • FBJ murine osteosarcoma viral v fos oncogene homolog antibody
      • FBJ Osteosarcoma Virus antibody
      • FOS antibody
      • FOS protein antibody
      • FOS_HUMAN antibody
      • G0 G1 switch regulatory protein 7 antibody
      • G0/G1 switch regulatory protein 7 antibody
      • G0S7 antibody
      • Oncogene FOS antibody
      • p55 antibody
      • proto oncogene c Fos antibody
      • Proto oncogene protein c fos antibody
      • Proto-oncogene c-Fos antibody
      • v fos FBJ murine osteosarcoma viral oncogene homolog antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [EPR20769] (ab214672)
      Immunocytochemistry/ Immunofluorescence - Anti-c-Fos antibody [EPR20769] (ab214672)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed. 0.1% Triton X-100 permeabilized serum treated and non-treated HeLa (human cervix adenocarcinoma epithelial cell) cells labeling c-Fos with ab214672 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing weakly nuclear staining on HeLa cells grown in serum free medium for 36 hours. Expression of c-Fos increased in HeLa cells grown in serum free medium for 36 hours followed by addition of 20% FBS for 2 hours.

      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.

    • Immunoprecipitation - Anti-c-Fos antibody [EPR20769] (ab214672)
      Immunoprecipitation - Anti-c-Fos antibody [EPR20769] (ab214672)

      c-Fos was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab214672 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab214672 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
      Lane 1: 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, 10 μg (Input).
      Lane 2: ab214672 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 ab214672 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: 1 second.

      The observed lower band is a proteasomal degradation fragment (PMID: 9737957).

    • Western blot - Anti-c-Fos antibody [EPR20769] (ab214672)
      Western blot - Anti-c-Fos antibody [EPR20769] (ab214672)
      All lanes : Anti-c-Fos antibody [EPR20769] (ab214672) at 1/1000 dilution

      Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum free medium for 36 hours, whole cell lysate
      Lane 2 : 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

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Developed using the ECL technique.

      Predicted band size: 40 kDa
      Observed band size: 55-60 kDa why is the actual band size different from the predicted?



      Exposure time: 15 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression of c-Fos is induced by the addition of 20%FBS (PMID: 24386331, PMID: 23300800, PMID: 25695333).

    • Western blot - Anti-c-Fos antibody [EPR20769] (ab214672)
      Western blot - Anti-c-Fos antibody [EPR20769] (ab214672)
      All lanes : Anti-c-Fos antibody [EPR20769] (ab214672) at 1/5000 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 (human T cell leukemia cell line from peripheral blood) 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

      Developed using the ECL technique.

      Predicted band size: 40 kDa
      Observed band size: 55-60 kDa why is the actual band size different from the predicted?



      Exposure time: 3 minutes.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      PMA treatment induces expression of c-Fos, as documented in the literature (PMID: 24386331, PMID: 23300800, PMID: 25695333).

    • Western blot - Anti-c-Fos antibody [EPR20769] (ab214672)
      Western blot - Anti-c-Fos antibody [EPR20769] (ab214672)
      All lanes : Anti-c-Fos antibody [EPR20769] (ab214672) at 1/5000 dilution

      Lane 1 : Untreated RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus grown in serum free medium overnight, whole cell lysate
      Lane 2 : RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) 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

      Developed using the ECL technique.

      Predicted band size: 40 kDa
      Observed band size: 55-60 kDa why is the actual band size different from the predicted?



      Exposure time: 1 minute.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      PMA treatment induces expression of c-Fos, as documented in the literature (PMID: 24386331, PMID: 23300800, PMID: 25695333).

    References

    This product has been referenced in:

    • Zhang Z  et al. METTL3-mediated N6-methyladenosine mRNA modification enhances long-term memory consolidation. Cell Res N/A:N/A (2018). WB ; Mouse . Read more (PubMed: 30297870) »
    See 1 Publication for this product

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