Key features and details
- Rabbit polyclonal to Cdc4 / Fbw7 / hSel 10
- Suitable for: WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Cdc4 / Fbw7 / hSel 10 antibody
See all Cdc4 / Fbw7 / hSel 10 primary antibodies
DescriptionRabbit polyclonal to Cdc4 / Fbw7 / hSel 10
SpecificityThe 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.
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic peptide corresponding to Human Cdc4/ Fbw7/ hSel 10 aa 650 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
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.
|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.|
RelevanceFbw7 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.
Cellular localizationCytoplasmic and Nuclear
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All lanes : Anti-Cdc4 / Fbw7 / hSel 10 antibody (ab12292) at 1/500 dilution
Lane 1 : HeLa nuclear lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : A6431 whole cell lysate
Lane 4 : Jurkat whole cell lysate
Lane 5 : HEK293 whole cell lysate
Lane 6 : HeLa nuclear lysate with Human Cdc4 / Fbw7 / hSel 10 peptide (ab12311) at 1 µg/ml
Lane 7 : HeLa whole cell lysate with Human Cdc4 / Fbw7 / hSel 10 peptide (ab12311) at 1 µg/ml
Lane 8 : A431 whole cell lysate with Human Cdc4 / Fbw7 / hSel 10 peptide (ab12311) at 1 µg/ml
Lane 9 : Jurkat whole cell lysate with Human Cdc4 / Fbw7 / hSel 10 peptide (ab12311) at 1 µg/ml
Lane 10 : HEK293 whole cell lysate with Human Cdc4 / Fbw7 / hSel 10 peptide (ab12311) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
All lanes : Rabbit Anti-Goat IgG H&L (HRP) (ab6741) at 1/5000 dilution
Predicted band size: 80 kDa
Exposure time: 10 seconds
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.
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.
All lanes : 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: 110,37 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutes
ab12292 has been referenced in 11 publications.
- Xu M et al. Interaction of YAP1 and mTOR promotes bladder cancer progression. Int J Oncol 56:232-242 (2020). PubMed: 31789387
- He J et al. Fbxw7 increases CCL2/7 in CX3CR1hi macrophages to promote intestinal inflammation. J Clin Invest 130:3877-3893 (2019). PubMed: 31246581
- Song Y et al. E3 ligase FBXW7 is critical for RIG-I stabilization during antiviral responses. Nat Commun 8:14654 (2017). Mouse . PubMed: 28287082
- Lv XB et al. Regulation of SOX10 stability via ubiquitination-mediated degradation by Fbxw7a modulates melanoma cell migration. Oncotarget 6:36370-82 (2015). PubMed: 26461473
- Balamurugan K et al. FBXW7a attenuates inflammatory signalling by downregulating C/EBPd and its target gene Tlr4. Nat Commun 4:1662 (2013). PubMed: 23575666