Overview

  • Product name
    Anti-eIF4E antibody [Y448]
    See all eIF4E primary antibodies
  • Description
    Rabbit monoclonal [Y448] to eIF4E
  • Host species
    Rabbit
  • Specificity
    The antibody detects a band on western blot of approximately 28 kDa.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human eIF4E aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control
    • 293 cell lysate, human breast carcinoma.
  • 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 ab33766 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
ICC/IF 1/250 - 1/500.
WB 1/500. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

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

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 [Y448] (ab33766) at 0.025 µg/ml

    Lane 1 : Raw264.7 (Mouse abelson murine leukemia virus-induced tumor) whole cell lysate
    Lane 2 : C6 (Rat glioma) 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



    Blocking and diluting buffer: 5% NFDM/TBST. 

    Exposure time - Lane 1: 160 seconds
                             Lane 2: 70 seconds 

  • Anti-eIF4E antibody [Y448] (ab33766) at 1/500 dilution + 293 cell lysate

    Predicted band size: 25 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?

  • This image shows human breast carcinoma stained with ab33766
  • Overlay histogram showing HEK293 cells stained with ab33766 (red line). The cells were fixed with 80% methanol (5 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. The cells were then incubated with the antibody (ab33766, 1µg/1x106 cells) 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 (1µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

  • ICC/IF image of ab33766 stained MCF7 cells. The cells were 4% formaldehyde (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 (ab33766, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

References

This product has been referenced in:
  • Li F  et al. Expression of 4E-BP1 and phospho-4E-BP1 correlates with the prognosis of patients with clear cell renal carcinoma. Cancer Manag Res 10:1553-1563 (2018). Read more (PubMed: 29942157) »
  • Tamarkin-Ben-Harush A  et al. Cap-proximal nucleotides via differential eIF4E binding and alternative promoter usage mediate translational response to energy stress. Elife 6:N/A (2017). Read more (PubMed: 28177284) »
See all 5 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cells (J1))
Total protein in input
600000 cells
Immuno-precipitation step
Other - Dynabeads
Specification
mouse embryonic stem cells (J1)

Abcam user community

Verified customer

Submitted Aug 03 2018

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

Abcam user community

Verified customer

Submitted Apr 10 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Fixation
Paraformaldehyde
Sample
Mouse Cell (T cells)
Specification
T cells
Gating Strategy
lymphocytes
Permeabilization
Yes - Phosflow PermBuffer III

Dr. Beata Zygmunt

Verified customer

Submitted Jun 25 2013

Answer

Thank you for your enquiry. In most cases, I would recommend using milk, because it works just as good as BSA and it is a bit cheaper. In the case of eIF4e though, because it is a phosphoprotein, you might get unspecific binding using milk because milk contains a phospoprotein called casein. This would lead to very high background, so in this case I would recommend BSA to block. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your enquiry. After contacting the originator of the antibody, I have updated the data sheet with the amino acid sequence which is NQEVANPEHYIKHPLQNR, which is amino acid residues 25-42, and it is specific for the N-terminus. I hope that this information helps. Please don't hesitate to ask us for help in the future.

Read More

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