Recombinant
RabMAb

Recombinant Anti-eIF4E antibody [Y448] (ab33766)

Reviews (3)Q&A (2)References (5)

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, IP, 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
    • WB: 293, HEK-293, and MCF7 cell lysates, Human brain tissue lysate IHC-P: human breast carcinoma, Human cervical carcinoma and Mouse stomach ICC/IF: RAW 264.7 cells FC: HEK-293 cells
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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.
IP 1/20.
WB 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa).

For unpurified use at 1/500

IHC-P 1/100.

See IHC antigen retrieval protocols.
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
    see all
  • 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

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4E antibody [Y448] (ab33766)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF4E antibody [Y448] (ab33766)

    Immunocytochemistry/ Immunofluorescence analysis of RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling eIF4E with Purified ab33766 at 1:500 dilution (0.3 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry - Anti-eIF4E antibody [Y448] (ab33766)
    Flow Cytometry - Anti-eIF4E antibody [Y448] (ab33766)

    Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling eIF4E with Purified ab33766 at 1:200 dilution (1 µg/ml) (red). Cells were fixed with 80% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling eIF4E with Purified ab33766 at 1:100 dilution (1.33 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunoprecipitation - Anti-eIF4E antibody [Y448] (ab33766)
    Immunoprecipitation - Anti-eIF4E antibody [Y448] (ab33766)

    ab33766 (purified) at 1:20 dilution (0.6µg) immunoprecipitating eIF4E in HEK-293 whole cell lysate.
    Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab33766 & HEK-293 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab33766 in HEK-293 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:10,000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse stomach tissue sections labeling eIF4E with Purified ab33766 at 1:100 dilution (1.33 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cervical carcinoma tissue sections labeling eIF4E with Purified ab33766 at 1:100 dilution (1.33 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Western blot - Anti-eIF4E antibody [Y448] (ab33766)
    Western blot - Anti-eIF4E antibody [Y448] (ab33766)
  • Western blot - Anti-eIF4E antibody [Y448] (ab33766)
    Western blot - Anti-eIF4E antibody [Y448] (ab33766)
  • Western blot - Anti-eIF4E antibody [Y448] (ab33766)
    Western blot - Anti-eIF4E antibody [Y448] (ab33766)
    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 

  • Western blot - Anti-eIF4E antibody [Y448] (ab33766)
    Western blot - Anti-eIF4E antibody [Y448] (ab33766)
    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?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E antibody [Y448] (ab33766)
    This image shows human breast carcinoma stained with ab33766
  • Flow Cytometry - Anti-eIF4E antibody [Y448] (ab33766)
    Flow Cytometry - Anti-eIF4E antibody [Y448] (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4E antibody [Y448] (ab33766)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF4E antibody [Y448] (ab33766)
    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

Publishing research using ab33766? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review

Filter by Application

Filter by Species

Filter by Ratings

1-3 of 3 Abreviews

Immunoprecipitation abreview for Anti-eIF4E antibody [Y448]

 Poor
Abreviews
abreview image
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)
Read More

Abcam user community

Verified customer

Submitted Aug 03 2018

Western blot abreview for Anti-eIF4E antibody [Y448]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Apr 10 2018

Flow Cytometry abreview for Anti-eIF4E antibody [Y448]

 Excellent
Abreviews
Application
Flow Cytometry
Fixation
Paraformaldehyde
Sample
Mouse Cell (T cells)
Specification
T cells
Gating Strategy
lymphocytes
Permeabilization
Yes - Phosflow PermBuffer III
Read More

Dr. Beata Zygmunt

Verified customer

Submitted Jun 25 2013

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up