Recombinant Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2151Y] to eIF4E (phospho S209)
- Suitable for: ICC/IF, WB, IP, IHC-P, Dot blot
- Reacts with: Mouse, Rat, Human, Pig
Related conjugates and formulations
Overview
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Product name
Anti-eIF4E (phospho S209) antibody [EP2151Y]
See all eIF4E primary antibodies -
Description
Rabbit monoclonal [EP2151Y] to eIF4E (phospho S209) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IP, IHC-P, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Pig -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: 293 cell lysate treated with alkaline phosphatase and HEK293 cell lysate treated with Dexamethasone. IHC-P: human breast carcinoma tissue. ICC/IF: HEK293 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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2151Y -
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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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76256 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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ICC/IF |
1/500.
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WB |
1/1000 - 1/100000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
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IP |
1/40 - 1/60.
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IHC-P | (1) |
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Dot blot |
1/1000.
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Notes |
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ICC/IF
1/500. |
WB
1/1000 - 1/100000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa). |
IP
1/40 - 1/60. |
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Dot blot
1/1000. |
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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All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/100000 dilution (purified)
Lane 1 : Untreated HEK293 whole cell lysate
Lane 2 : HEK293 cells treated with 10uM dexamethasone for 1 hour whole cell lysate
Lane 3 : HEK293 cells treated with 10uM dexamethasone for 1 hour whole cell lysate.The membrane was then incubated with alkaline phosphatase.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDa
Exposure time: 30 secondsBlocking buffer and concentration 2% BSA/TBST.
Diluting buffer and concentration 2% BSA/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling eIF4E with purified ab76256 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of untreated, 20% serum treated and 20% serum + LP treated NIH/3T3 cells labelling eIF4E (phospho S209) with ab76256 (left) and eIF4E with ab33766 (right) both at a dilution of 1/500.
Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
The image shows increased cytoplasmic staining after 20% serum treatment on NIH3T3 cells when compared with no serum treated cells. The LP treatment decreased the increased cytoplasmic staining caused by 20% serum.
ab33766 was used as a Pan control for ab76256. The results showed cytoplasmic staining on no serum, 20% serum and 20% serum +LP treated NIH3T3 cells.
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ab76256 (purified) at 1/40 immunoprecipitating eIF4E (phospho S209) in HEK293 whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1,500) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/1000 dilution (purified)
Lane 1 : Mouse spleen lysate
Lane 2 : Rat brain lysate
Lane 3 : Pig heart lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 25 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
Dot blot analysis of eIF4E (pS209) peptide (Lane 1) and eIF4E non-phospho peptide (Lane 2) labelling eIF4E (pS209) with purified ab76256 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Immunocytochemistry/Immunofluorescence analysis of serum starved HEK293 cells treated with CGP 57380 ab120365) labelling eIF4E (phospho S209) with unpurified ab32124 at 1/100. Decrease in eIF4E (phospho S209) expression correlates with increased concentration of CGP 57380, as described in literature.
The cells were incubated at 37°C for 1h in media containing different concentrations of ab120365 (CGP 57380) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with unpurified ab76256 was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/50000 dilution (purified)
Lane 1 : Untreated HEK293 cell lysate
Lane 2 : HEK293 treated with 10mM Dexamethasone 1 hour lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 25 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/50000 dilution (purified)
Lane 1 : Untreated 293 cell lysate
Lane 2 : 293 cell lysate treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Exposure time: 1 minuteBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/100000 dilution (purified)
Lane 1 : Untreated HEK293 cell lysate
Lane 2 : HEK293 cell lysate - treated with Dexamethasone
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Predicted band size: 25 kDaExposure time:
eIF4E pS209: 15 seconds.
eIF4E: 3 minutes.
Blocking and dilution buffer: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (49)
ab76256 has been referenced in 49 publications.
- Le Bras M et al. Translational Regulation by hnRNP H/F Is Essential for the Proliferation and Survival of Glioblastoma. Cancers (Basel) 14:N/A (2022). PubMed: 35267591
- Pham TN et al. Inhibition of MNKs promotes macrophage immunosuppressive phenotype to limit CD8+ T cell antitumor immunity. JCI Insight 7:N/A (2022). PubMed: 35380995
- Metz JB et al. High-throughput translational profiling with riboPLATE-seq. Sci Rep 12:5718 (2022). PubMed: 35383235
- Frosi Y et al. Engineering an autonomous VH domain to modulate intracellular pathways and to interrogate the eIF4F complex. Nat Commun 13:4854 (2022). PubMed: 35982046
- Shukla T et al. A Highly Selective MNK Inhibitor Rescues Deficits Associated with Fragile X Syndrome in Mice. Neurotherapeutics 18:624-639 (2021). PubMed: 33006091