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Anti-c-Fos (phospho T325) 抗体 (ab27793)

Reviews (1)Q&A (4)References (12)
Western blot - Anti-c-Fos (phospho T325) antibody (ab27793)
  • Immunocytochemistry/ Immunofluorescence - Anti-c-Fos (phospho T325) antibody (ab27793)
  • ChIP - Anti-c-Fos (phospho T325) antibody (ab27793)

Key features and details

  • Rabbit polyclonal to c-Fos (phospho T325)
  • Suitable for: ICC/IF, ChIP, WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-c-Fos (phospho T325) antibody
    c-Fos 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to c-Fos (phospho T325)

  • 由来種

    Rabbit

  • 特異性

    ab27793 recognises the cFos phosphorilated at threonine 325 form.

  • アプリケーション

    適用あり: ICC/IF, ChIP, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide corresponding to Human c-Fos (phospho T325).

  • ポジティブ･コントロール

    • WB: A431 cell lysate. ICC: HeLa cells. ChIP: A431 cells.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab27793の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ICC/IF
1/250.
ChIP
Use 1-3µg for 106 cells.
WB
1/1000. Predicted molecular weight: 41 kDa.
特記事項
ICC/IF
1/250.
ChIP
Use 1-3µg for 106 cells.
WB
1/1000. Predicted molecular weight: 41 kDa.

ターゲット情報

  • 機能

    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.

  • 配列類似性

    Belongs to the bZIP family. Fos subfamily.
    Contains 1 bZIP domain.

  • 翻訳後修飾

    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.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • 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

    画像

    • Western blot - Anti-c-Fos (phospho T325) antibody (ab27793)
      Western blot - Anti-c-Fos (phospho T325) antibody (ab27793)
      All lanes : Anti-c-Fos (phospho T325) antibody (ab27793) at 1/1000 dilution (diluted in a 3% Milk TBST buffer.)

      Lane 1 : Non EGF treated A431 cell lysate
      Lane 2 : EGF treated A431 cell lysate
      Lane 3 : EGF treated A431 cell lysate treated with Lambda phosphatase

      Secondary
      All lanes : goat F(ab’)2 anti rabbit IgG HRP conjugate

      Predicted band size: 41 kDa
      Observed band size: 58 kDa why is the actual band size different from the predicted?



      The figure shows that the phosphorylation of cFos on threonine 325 is induced by EGF treatment and that Lambda phosphatase treatment eliminates the signal, thereby demonstrating the phospho specificity of ab27793.
    • Immunocytochemistry/ Immunofluorescence - Anti-c-Fos (phospho T325) antibody (ab27793)
      Immunocytochemistry/ Immunofluorescence - Anti-c-Fos (phospho T325) antibody (ab27793)

      Immunofluorescence analysis of c-Fos (phospho T325) was done on 70% confluent log phase HeLa cell treated with 200 nM of PMA for 20 minutes. The cell were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phosc-Fos (phospho T325)Rabbit Polyclonal Antibody (ab27793) at 2 ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1/2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin. Panel d is a merged image showing nuclear and cytoplasmic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

    • ChIP - Anti-c-Fos (phospho T325) antibody (ab27793)
      ChIP - Anti-c-Fos (phospho T325) antibody (ab27793)

      Chromatin Immunoprecipitation (ChIP) was performed using Anti-c-Fos (phospho T325) antibody (ab27793) 3 ug on sheared chromatin from 2 million A431 cells treated with EGF (200ng/ml), for 30 minutes. Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system with optimized PCR primer pairs for the promoter of active IL-6, CDKN1A gene, used as positive control target, and the SAT2, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

    参考文献 (12)

    ab27793 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab27793 は 12 報の論文で使用されています。

    • Usuki S  et al. Konjac Ceramide (kCer)-Mediated Signal Transduction of the Sema3A Pathway Promotes HaCaT Keratinocyte Differentiation. Biology (Basel) 11:N/A (2022). PubMed: 35053118
    • Cao B  et al. Extract of Corallodiscus flabellata attenuates renal fibrosis in SAMP8 mice via the Wnt/β-catenin/RAS signaling pathway. BMC Complement Med Ther 22:52 (2022). PubMed: 35227255
    • Dou C  et al. Sialylation of TLR2 initiates osteoclast fusion. Bone Res 10:24 (2022). PubMed: 35232979
    • Gao W  et al. Psoralen inhibits the inflammatory response and mucus production in allergic rhinitis by inhibiting the activator protein 1 pathway and the downstream expression of cystatin‑SN. Mol Med Rep 24:N/A (2021). PubMed: 34278468
    • Binato R  et al. NRIP1 is activated by C-JUN/C-FOS and activates the expression of PGR, ESR1 and CCND1 in luminal A breast cancer. Sci Rep 11:21159 (2021). PubMed: 34707101
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-c-Fos (phospho T325) antibody

     Inconclusive
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Chicken Cell lysate - whole cell (Chicken thrombocyte)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Specification
    Chicken thrombocyte
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Mrs. Katie Elliott

    Verified customer

    投稿 Apr 25 2017

    Question

    The order no was XXX and it was placed on 22.03.12.

    Read More
    Answer

    Thank you for contacting us.

    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Question

    I have tried the suggestions that you made but am still not satisfied with the antibody. I would appreciate it if you could arrange a refund of this product.

    Read More
    Answer

    Thank you for your reply.


    I am sorry to hear that my suggestions did not improve your results with this antibody.I am happy to arrange for a refund. Would you please provide your order number for this product?


    I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

    Read More

    Question

    I recently purchased the antibody ab27793 - phospho c-Fos T325 but it has not worked at all well. I have tried it on both mouse embryonic fibroblast and motor neuron lysates. Both cell types had been stimulated with growth factors and so there should have been plenty of phosphorylated protein to detect. I blocked the membranes for 1 hour in PBS-T 0.2% with 5% non-fat milk. I then incubated with the primary antibody (1:1,000) over night at 4 C in PBS-T 0.2% with the addition of 5% BSA. The membrane was incubated with an HRP conjugated rabbit secondary antibody, in PBS-T 0.2% and 5% non-fat milk for 45 min. In between the primary and secondary antibodies and also post secondary antibody the membranes were washes in PBS-T 0.2% for 15 min x1 followed by 5 min x3, although I did try longer washing steps also. There are multiple bands on western blots, some of which vary between membranes. There was also a lot of background signal detected making it very difficult to distinguish what was going on.
    Any advice would be appreciated and/or a refund arranged.

    Read More
    Answer

    Thank you for contacting Abcam regarding ab27793.


    I am sorry that you have been experiencing difficulties with this antibody in WB. I have reviewed the protocol information you provided and would like to make some suggestions to improve your results.


    When using a phospho-specific antibody it is highly advised against using milk as a blocking reagent or diluent. I would recommend switching to 5% BSA in PBST instead. Also switching to TBST often decreases background. I would suggest the following:


    How much sample are you loading? I would recommend loading 50-75ug total protein in each lane. Then, block for at least 1 hr at room temperature in 5% BSA in TBST. Incubate with the primary antibody (also diluted in 5% BSA in TBST) for 1 hour at room temperature rather than overnight. Test 1:1000, 1:2000, and 1:4000. Perform washes with TBST. Incubate the secondary antibody diluted in 5% BSA (dilute according to manufacturers instructions). It may be necessary to titrate the secondary antibody concentration as well. Wash and develop as you normally would.


    I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or if these suggestions do not improve your results with this antibody and I will be happy to provide a replacement or refund per our Abpromise guarantee.

    Read More

    Question

    I notice that you have phospho c-fos antibodies available. Have these antibodies been used for immunofluorescence in tissues?

    Read More
    Answer

    Thank you for your enquiry. Ab27793 has not yet been tested for application in IF (only Western blotting so far)but ab17933 - Rabbit polyclonal to c-Fos (phospho T232) - has been tested in IF. There is an image on the online datasheet with more more information. Please contact us again if you have any additional questions.

    Read More

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