Anti-Cdc4 / Fbw7 / hSel 10抗体(ab12292)

概述

  • 产品名称Anti-Cdc4 / Fbw7 / hSel 10抗体
    参阅全部 Cdc4 / Fbw7 / hSel 10 一抗
  • 描述
    兔多克隆抗体to Cdc4 / Fbw7 / hSel 10
  • 特异性The immunising sequence is present in all 4 of the human isoforms. We have tested the antibody on Hela, A431, Jurkat and 293 cells, but to date have seen no clear bands at the expected molecular weights of 79.7kDa, 70.3kDa, 69.7kDa and 66.2 kDa. According to the literature, the alpha and gamma isoforms are localized to the nucleus and the beta isoform is localized to cytoplasm. The ICC/IF data that we have generated in-house shows both nuclear and cytoplasmic staining and as such we believe ab12292 is detecting Cdc4/Fbw7/hSel10, however we would appreciate any further confirmation of this.
  • 经测试应用适用于: WB, ICC/IF, IPmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Human Cdc4/ Fbw7/ hSel 10.

    (Peptide available as ab12311.)

  • 阳性对照
    • This antibody gave a positive signal in Hela and Jurkat Whole Cell and Nuclear lysates and in A431 and HEK293 Whole Cell Lysates

性能

应用

Our Abpromise guarantee covers the use of ab12292 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/500. Detects a band of approximately 45,110 kDa (predicted molecular weight: 80 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml. PubMed: 20388706

靶标

  • 相关性Fbw7 is a member of the F box protein family which are characterized by an approximately 40 amino acid motif, the F box. The F box proteins constitute one of the four subunits of ubiquitin protein ligase complex called SCFs (SKP1-cullin-F box), which function in phosphorylation-dependent ubiquitination. The F box proteins are divided into 3 classes: Fbws containing WD40 domains, Fbls containing leucine rich repeats, and Fbxs containing either different protein-protein interaction modules or no recognizable motifs. Fbw7 belongs to the Fbws class; in addition to an F box, this protein contains 7 tandem WD40 repeats. It binds directly to cyclin E and probably targets cyclin E for ubiquitin mediated degradation. Mutations of this gene are detected in ovarian and breast cancer cell lines. Alternative splicing of this gene generates 2 transcript variants diverging at the 5' termini.
  • 细胞定位Cytoplasmic and Nuclear
  • 数据库链接
  • 别名
    • AGO antibody
    • Archipelago Drosophila homolog of antibody
    • Archipelago homolog antibody
    • CDC 4 antibody
    • CDC4 antibody
    • DKFZp686F23254 antibody
    • F box and WD 40 domain protein 7 antibody
    • F box protein FBW7 antibody
    • F box protein FBX30 antibody
    • F box protein SEL 10 antibody
    • F box/WD repeat protein 7 antibody
    • FBW 7 antibody
    • FBW6 antibody
    • FBW7 antibody
    • FBX 30 antibody
    • FBXO30 antibody
    • FBXW 6 antibody
    • FBXW 7 antibody
    • FBXW7 antibody
    • hAgo antibody
    • hCdc4 antibody
    • Homolog of C elegans sel 10 antibody
    • SEL10 antibody
    see all

Anti-Cdc4 / Fbw7 / hSel 10 antibody 图像



  • Predicted band size : 80 kDa

    Western blot using ab12292 at 1/500.

    Lane 1: HeLa nuclear
    Lane 2: HeLa whole cell
    Lane 3: A431 whole cell
    Lane 4: Jurkat whole cell
    Lane 5: HEK293 whole cell
    Lane 6: HeLa nuclear + blocking peptide (ab12311)
    Lane 7: HeLa whole cell + blocking peptide (ab12311)
    Lane 8: A431 whole cell + blocking peptide (ab12311)
    Lane 9: Jurkat whole cell + blocking peptide (ab12311)
    Lane 10: HEK293 whole cell + blocking peptide (ab12311)

    Secondary: Goat polyclonal to Rabbit IgG HRP ab6741 (1/5000)
    Exposure time: 10 seconds.
    20µg of cell lysate per lane
    Blocking peptide used at 1µg/ml.

    Expected band size at 80kDa.
    Additional bands seen at 45, 55 and 150 kDa. We are uncertain as to the identity of these bands.

     

  • ab12292 (1/2000) detecting Cdc4 in Hela cells (green). Cells were fixed in methanol, permeabilised with 0.5% Triton X100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • ICC/IF image of ab12292 stained HeLa cells. The cells were 4% PFA 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 (ab12292, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2 and MCF7 cells at 5µg/ml.
  • All lanes : Anti-Cdc4 / Fbw7 / hSel 10 antibody (ab12292) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 80 kDa
    Observed band size : 37,110 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

Anti-Cdc4 / Fbw7 / hSel 10 antibody (ab12292)参考文献

This product has been referenced in:
  • Balamurugan K  et al. FBXW7a attenuates inflammatory signalling by downregulating C/EBPd and its target gene Tlr4. Nat Commun 4:1662 (2013). Read more (PubMed: 23575666) »
  • Biswas M  et al. Glycogen synthase kinase 3 regulates expression of nuclear factor-erythroid-2 related transcription factor-1 (Nrf1) and inhibits pro-survival function of Nrf1. Exp Cell Res 319:1922-31 (2013). Read more (PubMed: 23623971) »

See all 6 Publications for this product

Product Wall

Thank you very much for contacting us with your question.

This antibody was tested with a 1 hour incubation at room temperature, but an overnight incubation will be fine as well if that is more convenient for you. The membrane was blocked wit...

Read More

Thank you for contacting us.
Due to our large number of products and limited storage caopacity we are unable to provide any trial or sample sizes. The upper WB image does show nicely the epxted bands in Hela, HEK293, and Jurkat cells with and with...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell (SPhi992 strain)
Loading amount 50 µg
Specification SPhi992 strain
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Odyssey Blocking Agent (Lycor system) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 18 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (MPRO (mouse promyelocytic) cells)
Loading amount 50 µg
Specification MPRO (mouse promyelocytic) cells
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Odyssey Blocking Agent (Lycor system) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 18 2010

Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 50 µg
Specification HEK293
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Odyssey Blocking Agent (Lycor system) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 18 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (kidney)
Specification kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 05 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

提交于 Oct 30 2007

Thank you for your enquiry. I can confirm that Fbw7 has several known isoforms. The better known isoforms 1 and 2 have molecular weights of 69 and 110kDa. Further information on the isoforms can be found via the SwissProt protein database link on t...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"