Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y449] to eIF4E
- Suitable for: WB, IHC-P, ICC, Flow Cyt, IP
- Reacts with: Mouse, Rat, Human
Product nameAnti-eIF4E antibody [Y449]
See all eIF4E primary antibodies
DescriptionRabbit monoclonal [Y449] to eIF4E
Tested applicationsSuitable for: WB, IHC-P, ICC, Flow Cyt, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human eIF4E aa 150-250. The exact sequence is proprietary.
- WB: HEK-293, HepG2, NIH/3T3, RAW 264.7, and C6 cell lysates; IHC-P: Human ovarian cancer tissue. Mouse and rat testis tissues; ICC/IF: RAW 264.7 cells; Flow Cyt: HEK293 cells; IP: NIH/3T3 cell lysate.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|WB||1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa).
For unpurified use at 1/5000 - 1/20000.
|IHC-P||1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionIts 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 similaritiesBelongs to the eukaryotic initiation factor 4E family.
modificationsPhosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
- Information by UniProt
- AUTS19 antibody
- CBP antibody
- eIF 4E antibody
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.
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.
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.
Immunocytochemistry/ Immunofluorescence analysis of RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling eIF4E with Purified ab33768 at 1/50 (10 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of HEK-293 (Human embryonickidney epithelial cell) cells labeling eIF4E with Purified ab33768 at 1/50 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% 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).
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.
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.
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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab33768 has been referenced in 1 publication.
- 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). PubMed: 28610844