Anti-c-Jun (phospho T91 + T93) antibody [C-J 4C4] (ab13671)

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Key features and details

  • Mouse monoclonal [C-J 4C4] to c-Jun (phospho T91 + T93)
  • Suitable for: IHC-P, WB, ICC/IF
  • Reacts with: Rat, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-c-Jun (phospho T91 + T93) antibody [C-J 4C4]
    See all c-Jun primary antibodies

  • Description

    Mouse monoclonal [C-J 4C4] to c-Jun (phospho T91 + T93)

  • Host species

    Mouse

  • Tested applications

    Suitable for: IHC-P, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    c-Jun protein phosphorylated at T91 and T93.

  • Positive control

    • This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.

Properties

Applications

Our Abpromise guarantee covers the use of ab13671 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
IHC-P Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function

    Transcription factor that recognizes and binds to the enhancer heptamer motif 5'-TGA[CG]TCA-3'. Promotes activity of NR5A1 when phosphorylated by HIPK3 leading to increased steroidogenic gene expression upon cAMP signaling pathway stimulation. Involved in activated KRAS-mediated transcriptional activation of USP28 in colorectal cancer (CRC) cells (PubMed:24623306). Binds to the USP28 promoter in colorectal cancer (CRC) cells (PubMed:24623306).

  • Sequence similarities

    Belongs to the bZIP family. Jun subfamily.
    Contains 1 bZIP (basic-leucine zipper) domain.

  • Post-translational
    modifications

    Ubiquitinated by the SCF(FBXW7), leading to its degradation. Ubiquitination takes place following phosphorylation, that promotes interaction with FBXW7.
    Phosphorylated by CaMK4 and PRKDC; phosphorylation enhances the transcriptional activity. Phosphorylated by HIPK3. Phosphorylated by DYRK2 at Ser-243; this primes the protein for subsequent phosphorylation by GSK3B at Thr-239. Phosphorylated at Thr-239, Ser-243 and Ser-249 by GSK3B; phosphorylation reduces its ability to bind DNA. Phosphorylated by PAK2 at Thr-2, Thr-8, Thr-89, Thr-93 and Thr-286 thereby promoting JUN-mediated cell proliferation and transformation. Phosphorylated by PLK3 following hypoxia or UV irradiation, leading to increase DNA-binding activity.
    Acetylated at Lys-271 by EP300.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Activator protein 1 antibody
      • AP 1 antibody
      • AP-1 antibody
      • AP1 antibody
      • cJun antibody
      • Enhancer Binding Protein AP1 antibody
      • Jun Activation Domain Binding Protein antibody
      • JUN antibody
      • Jun oncogene antibody
      • JUN protein antibody
      • Jun proto oncogene antibody
      • JUN_HUMAN antibody
      • JUNC antibody
      • Oncogene JUN antibody
      • p39 antibody
      • Proto oncogene c jun antibody
      • Proto oncogene cJun antibody
      • Proto-oncogene c-jun antibody
      • Transcription Factor AP 1 antibody
      • Transcription factor AP-1 antibody
      • Transcription Factor AP1 antibody
      • V jun avian sarcoma virus 17 oncogene homolog antibody
      • V jun sarcoma virus 17 oncogene homolog (avian) antibody
      • V jun sarcoma virus 17 oncogene homolog antibody
      • V-jun avian sarcoma virus 17 oncogene homolog antibody
      • vJun Avian Sarcoma Virus 17 Oncogene Homolog antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho T91 + T93) antibody [C-J 4C4] (ab13671)
      Immunocytochemistry/ Immunofluorescence - Anti-c-Jun (phospho T91 + T93) antibody [C-J 4C4] (ab13671)
      ICC/IF image of ab13617 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13617, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho T91 + T93) antibody [C-J 4C4] (ab13671)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun (phospho T91 + T93) antibody [C-J 4C4] (ab13671)

      IHC image of c-Jun (phospho T91 + T93) staining in Human normal lung formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab13671, 5µg/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.

      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.

    References

    Publishing research using ab13671? Please let us know so that we can cite the reference in this datasheet.

    ab13671 has been referenced in 6 publications.

    • Hu G  et al. MicroRNA-145 attenuates TNF-a-driven cartilage matrix degradation in osteoarthritis via direct suppression of MKK4. Cell Death Dis 8:e3140 (2017). IHC-P ; Human . PubMed: 29072705
    • Kudo K  et al. Inhibition of Gli1 results in altered c-Jun activation, inhibition of cisplatin-induced upregulation of ERCC1, XPD and XRCC1, and inhibition of platinum-DNA adduct repair. Oncogene 31:4718-24 (2012). Human . PubMed: 22266871
    • Madeo A  et al. c-Jun activation is required for 4-hydroxytamoxifen-induced cell death in breast cancer cells. Oncogene 29:978-91 (2010). WB ; Human . PubMed: 19935718
    • Vinciguerra M  et al. Negative charged threonine 95 of c-Jun is essential for c-Jun N-terminal kinase-dependent phosphorylation of threonine 91/93 and stress-induced c-Jun biological activity. Int J Biochem Cell Biol 40:307-16 (2008). PubMed: 17920329
    • Hartz AM  et al. Diesel exhaust particles induce oxidative stress, proinflammatory signaling, and P-glycoprotein up-regulation at the blood-brain barrier. FASEB J : (2008). PubMed: 18474546
    • Nateri AS  et al. The ubiquitin ligase SCFFbw7 antagonizes apoptotic JNK signaling. Science 303:1374-8 (2004). PubMed: 14739463

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    I recently purchased your Anti-c-Jun Antibody, ab13671, but I am having difficulties getting it to work. Could you suggest some lysates for me to use? Currently, I have been using it for Western Blots, against Jurkat, HeLa and 3T3 lysates, which are treated with Anisomycin. I have not gotten any signals at all. My current procedures are as follows: I load 20-30 ug of each lysate on a 12 % gel, followed by transfer onto nitrocellulose and/or PVDF membrane (I've tried both unsucessfully). I block for an hour, then incubate with your antibody at 5 ug/mL for another hour. I use a Goat anti-Mouse IgG:AP secondary for an hour. I've repeated this numerous times and I still have yet to see a signal. Any help would be much appreciated.

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    Answer

    Thank you for your enquiry and your patience. I am very sorry to hear that you are having problem with this antibody. I would like to make the following suggestions/comments: 1. c-Jun is a nuclear protein, so try to prepare nuclear lysate rather than whole cell lysate. Make sure the lysis buffer contains a cocktail of different proteinase inhibitors in order to prevent the degradation of the target protein. 2. Incubate the membrane overnight at 4oC and check if you have stronger signal or not. 3. If you are using an antibody that is specific to a phosphorylated epitope, do not use milk to block. Kinases from the milk will non-specifically phosphorylate proteins in your samples. Application of casein or milk may lead to decrease or loss of signal intensity due to reaction of the primary antibody with the blocking/diluting agent. We recommend replacing milk by 5% BSA. It is also important to block overnight. Finally, please take a look at this specific publication, you may find some useful information: Nateri AS et al. The ubiquitin ligase SCFFbw7 antagonizes apoptotic JNK signaling. Science 303:1374-8 (2004). PubMed: 14739463 I hope this will be useful for you. Should you still have any problem, then please do not hesitate to contact us again.

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    Question

    I have a question regarding your c-Jun phospho antibody. Can I use anisomycin stimulated HeLa cell lysate as positive control? Or do you have a positive cell lysate available in your catalog?

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    Answer

    Thank you very much for your enquiry and patience. I have contacted the originator of this antibody for more information regarding what is recommended as a positive control and am waiting for them to get back to me. As soon as I get an answer, I will contact you. In the meantime I suggest referring the following publication which used this antibody: Nateri AS et al. The ubiquitin ligase SCFFbw7 antagonizes apoptotic JNK signaling. Science 303:1374-8 (2004). PubMed: 14739463

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