Recombinant 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: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human eIF4E aa 1-100 (N terminal). The exact sequence is proprietary.
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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 (intra): HEK-293 cells
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y448 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
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 |
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Flow Cyt (Intra) |
Use 1µg for 106 cells.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/250 - 1/500.
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IP | (1) |
1/20.
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WB | (6) |
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.
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Notes |
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Flow Cyt (Intra)
Use 1µg for 106 cells. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
Target
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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
modificationsPhosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex. - Information by UniProt
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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
see all -
Alternative names
- AUTS19 antibody
- CBP antibody
- eIF 4E antibody
see all
Images
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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.
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Intracellular Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling eIF4E with Purified ab33766 at 1/200 dilution (1µg/ml) (red). Cells were fixed with 80% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 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).
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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.
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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) 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.
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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.
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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 kDaBlocking and dilutin buffer and concentration: 5% NFDM/TBST
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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 kDaBlocking and dilutin buffer and concentration: 5% NFDM/TBST
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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 kDaBlocking 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
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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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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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.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (13)
ab33766 has been referenced in 13 publications.
- Shad BJ et al. Daily Myofibrillar Protein Synthesis Rates in Response to Low- and High-Frequency Resistance Exercise Training in Healthy, Young Men. Int J Sport Nutr Exerc Metab 31:209-216 (2021). PubMed: 33601335
- Zhang Y et al. Inhibition of miR-15a-5p Promotes the Chemoresistance to Pirarubicin in Hepatocellular Carcinoma via Targeting eIF4E. Comput Math Methods Med 2021:6468405 (2021). PubMed: 34812269
- 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
- Lemaire J et al. Zinc interference with Cd-induced hormetic effect in differentiated Caco-2 cells: Evidence for inhibition downstream ERK activation. J Biochem Mol Toxicol 34:e22437 (2020). PubMed: 31860779
- Xu M et al. Interaction of YAP1 and mTOR promotes bladder cancer progression. Int J Oncol 56:232-242 (2020). PubMed: 31789387