Name: Anti-eIF4E antibody [Y448]
Brand: RabMAb
SKU: ab33766
Rating: 4 (3 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y448] to eIF4E
Suitable for: ICC/IF, IP, WB, IHC-P, Flow Cyt
Reacts with: Mouse, Rat, Human

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 stomachICC/IF: RAW 264.7 cells Flow Cyt: HEK-293 cells
General notes
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
For more information see here.
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y448
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HEK-293 whole cell lysate (ab7902)
Recombinant Protein
- Recombinant Human eIF4E protein (ab56276)

The Abpromise guarantee

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)	1/20.
WB	(1)	1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 25 kDa). For unpurified use at 1/500
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
Flow Cyt	(1)	Use 1µg for 10⁶ cells. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. 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.

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.
Target information above from: UniProt accession P06730 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 1977 Human
- Entrez Gene: 13684 Mouse
- Entrez Gene: 117045 Rat
- Omim: 133440 Human
- SwissProt: P06730 Human
- SwissProt: P63073 Mouse
- SwissProt: P63074 Rat
- Unigene: 249718 Human
- Unigene: 3941 Mouse
- Unigene: 11275 Rat
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

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 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) 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)

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) 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) 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)

Anti-eIF4E antibody [Y448] (ab33766) at 1/1000 dilution (Purified) + Human brain lysates at 15 µg

Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 25 kDa
Observed band size: 25 kDa

Blocking and dilutin buffer and concentration: 5% NFDM/TBST
Western blot - Anti-eIF4E antibody [Y448] (ab33766)

All lanes : Anti-eIF4E antibody [Y448] (ab33766) at 1/5000 dilution

Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates

Lysates/proteins at 15 µg per lane.

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

Predicted band size: 25 kDa
Observed band size: 25 kDa

Blocking and dilutin buffer and concentration: 5% NFDM/TBST
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)

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)

This image shows human breast carcinoma stained with 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)

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.
Anti-eIF4E antibody [Y448] (ab33766)

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.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

ab33766 has been referenced in 10 publications.

Xu M et al. Interaction of YAP1 and mTOR promotes bladder cancer progression. Int J Oncol 56:232-242 (2020). PubMed: 31789387
Jin X et al. Cinobufagin Triggers Defects in Spindle Formation and Cap-Dependent Translation in Liver Cancer Cells by Inhibiting the AURKA-mTOR-eIF4E Axis. Am J Chin Med 48:651-678 (2020). PubMed: 32349518
Zhang Y et al. Non-genomic mechanisms mediate androgen-induced PSD95 expression. Aging (Albany NY) 11:2281-2294 (2019). PubMed: 31005955
Haimov O et al. Dynamic Interaction of Eukaryotic Initiation Factor 4G1 (eIF4G1) with eIF4E and eIF1 Underlies Scanning-Dependent and -Independent Translation. Mol Cell Biol 38:N/A (2018). PubMed: 29987188
Li Z et al. Epithelial mesenchymal transition induced by the CXCL9/CXCR3 axis through AKT activation promotes invasion and metastasis in tongue squamous cell carcinoma. Oncol Rep 39:1356-1368 (2018). PubMed: 29286143
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). 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). PubMed: 28177284
Qiao Y et al. High Glucose Stimulates Tumorigenesis in Hepatocellular Carcinoma Cells Through AGER-Dependent O-GlcNAcylation of c-Jun. Diabetes 65:619-32 (2016). PubMed: 26825459
Wang J et al. Doxorubicin induces apoptosis by targeting Madcam1 and AKT and inhibiting protein translation initiation in hepatocellular carcinoma cells. Oncotarget 6:24075-91 (2015). IP . PubMed: 26124182
Wang M et al. WWOX suppresses cell growth and induces cell apoptosis via inhibition of P38 nuclear translocation in cholangiocarcinoma. Cell Physiol Biochem 34:1711-22 (2014). PubMed: 25502636

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1-3 of 3 Abreviews

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

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

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

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (10)

Customer reviews and Q&As

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Immunoprecipitation abreview for Anti-eIF4E antibody [Y448]

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Post-translational
modifications