Recombinant
RabMAb

Recombinant Anti-eIF4E antibody [Y449] (ab33768)

Rabbit recombinant monoclonal eIF4E antibody [Y449]. Validated in WB, IP, IHC, Flow Cyt and tested in Mouse, Rat, Human. Cited in 1 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name
    Anti-eIF4E antibody [Y449]
    See all eIF4E primary antibodies
  • Description
    Rabbit monoclonal [Y449] to eIF4E
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
    Unsuitable for: ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human eIF4E aa 150-250. The exact sequence is proprietary.

  • Positive control
    • WB: HEK-293, HepG2, NIH/3T3, RAW 264.7, PC-12, and C6 cell lysates. IHC-P: Human breast carcinoma, human ovarian cancer and mouse and rat testis tissues. Flow Cyt: HEK293 cells. IP: NIH/3T3 cell lysate.
  • 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 ab33768 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
WB 1/5000 - 1/20000. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/30.
  • Application notes
    Is unsuitable for ICC.
  • Target

    • Function
      Its translation stimulation activity is repressed by binding to the complex CYFIP1-FMR1 (By similarity). Recognizes and binds the 7-methylguanosine-containing mRNA cap during an early step in the initiation of protein synthesis and facilitates ribosome binding by inducing the unwinding of the mRNAs secondary structures. Component of the CYFIP1-EIF4E-FMR1 complex which binds to the mRNA cap and mediates translational repression. In the CYFIP1-EIF4E-FMR1 complex this subunit mediates the binding to the mRNA cap.
    • Sequence similarities
      Belongs to the eukaryotic initiation factor 4E family.
    • Post-translational
      modifications
      Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
    • Information by UniProt
    • Database links
    • Alternative names
      • AUTS19 antibody
      • CBP antibody
      • eIF 4E antibody
      • eIF 4F 25 kDa subunit antibody
      • EIF 4F antibody
      • eIF-4E antibody
      • eIF-4F 25 kDa subunit antibody
      • eIF4E antibody
      • EIF4E1 antibody
      • EIF4EL1 antibody
      • EIF4F antibody
      • Eukaryotic translation initiation factor 4 E antibody
      • Eukaryotic translation initiation factor 4E antibody
      • Eukaryotic translation initiation factor 4E like 1 antibody
      • IF4E_HUMAN antibody
      • Messanger RNA Cap Binding Protein eIF 4E antibody
      • MGC111573 antibody
      • mRNA cap binding protein antibody
      • mRNA cap-binding protein antibody
      see all

    Images

    • All lanes : Anti-eIF4E antibody [Y449] (ab33768) at 1/1000 dilution

      Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
      Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
      Lane 4 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 25 kDa
      Observed band size: 28 kDa
      why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time:
      Lane 1: 20 seconds.
      Lanes 2-5: 8 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian cancer tissue labelling eIF4E with ab33768 at a dilution of 1/500. Antigen retrieval was performed ab93684, Tris/EDTA buffer, pH 9. A ready to use goat anti-rabbit IgG H&L (HRP Polymer) was used as the secondary antibody. Counter stained with Hematoxylin.

    • eIF4E was immunoprecipitated from 10 µg NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate with ab33768 at a 1/30 dilution. Western blot was performed from the immunoprecipitate using ab33768 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg (Input).
      Lane 2: ab33768 IP in NIH/3T3 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab33768 in NIH/3T3 whole cell lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Overlay histogram showing HEK293 cells stained with ab33768 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33768, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue labelling eIF4E with ab33768 at a dilution of 1/500. Antigen retrieval was performed ab93684, Tris/EDTA buffer, pH 9. A ready to use goat anti-rabbit IgG H&L (HRP Polymer) was used as the secondary antibody. Counter stained with Hematoxylin.

    • All lanes : Anti-eIF4E antibody [Y449] (ab33768) at 1/500 dilution

      Lane 1 : C6 (Rat glioma) whole cell lysate
      Lane 2 : Raw264.7 (Mouse abelson murine leukemia virus-induced tumor) whole cell lysate
      Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
      Lane 4 : NIH/3T3 (Mouse embryo) whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 25 kDa
      Observed band size: 28 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue labelling eIF4E with ab33768 at a dilution of 1/500. Antigen retrieval was performed ab93684, Tris/EDTA buffer, pH 9. A ready to use goat anti-rabbit IgG H&L (HRP Polymer) was used as the secondary antibody. Counter stained with Hematoxylin.

    • Ab33768, at 1/250 dilution, staining eIF4E in paraffin embedded human breast carcinoma tissue sections by Immunohistochemistry.

    References

    This product has been referenced in:
    • Oblinger JL  et al. Overexpression of eIF4F components in meningiomas and suppression of meningioma cell growth by inhibiting translation initiation. Exp Neurol 299:299-307 (2018). Read more (PubMed: 28610844) »
    See 1 Publication for this product

    Customer reviews and Q&As

    Filter by Application

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    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse embryonic stem cells (J1))
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    mouse embryonic stem cells (J1)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

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    Verified customer

    Submitted Apr 10 2018

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