Rabbit Recombinant Monoclonal eIF4E antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Rat, Human samples. Cited in 4 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
ICC/IF | IP | WB | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|
Human | Expected | Expected | Tested | Tested | Tested |
Mouse | Tested | Tested | Tested | Expected | Tested |
Rat | Expected | Expected | Tested | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes For unpurified use at 1/5000 - 1/20000. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/1000 | Notes For unpurified use at 1/5000 - 1/20000. |
Species Human | Dilution info 1/1000 | Notes For unpurified use at 1/5000 - 1/20000. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Acts in the cytoplasm to initiate and regulate protein synthesis and is required in the nucleus for export of a subset of mRNAs from the nucleus to the cytoplasm which promotes processes such as RNA capping, processing and splicing (PubMed:11606200, PubMed:22578813, PubMed:22684010, PubMed:24335285, PubMed:29987188). Component of the protein complex eIF4F, which is involved in the recognition of the mRNA cap, ATP-dependent unwinding of 5'-terminal secondary structure and recruitment of mRNA to the ribosome (By similarity). This protein recognizes and binds the 7-methylguanosine (m7G)-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 (PubMed:16271312, PubMed:22578813). Together with EIF4G1, antagonizes the scanning promoted by EIF1-EIF4G1 and is required for TISU translation, a process where the TISU element recognition makes scanning unnecessary (PubMed:29987188). In addition to its role in translation initiation, also acts as a regulator of translation and stability in the cytoplasm (PubMed:24335285). Component of the CYFIP1-EIF4E-FMR1 complex which binds to the mRNA cap and mediates translational repression: in the complex, EIF4E mediates the binding to the mRNA cap (By similarity). Component of a multiprotein complex that sequesters and represses translation of proneurogenic factors during neurogenesis (By similarity). In P-bodies, component of a complex that mediates the storage of translationally inactive mRNAs in the cytoplasm and prevents their degradation (PubMed:24335285). May play an important role in spermatogenesis through translational regulation of stage-specific mRNAs during germ cell development (By similarity). As well as its roles in translation, also involved in mRNA nucleocytoplasmic transport (By similarity). Its role in mRNA export from the nucleus to the cytoplasm relies on its ability to bind the m7G cap of RNAs and on the presence of the 50-nucleotide EIF4E sensitivity element (4ESE) in the 3'UTR of sensitive transcripts (By similarity). Interaction with the 4ESE is mediated by LRPPRC which binds simultaneously to both EIF4E and the 4ESE, thereby acting as a platform for assembly for the RNA export complex (By similarity). EIF4E-dependent mRNA export is independent of ongoing protein or RNA synthesis and is also NFX1-independent but is XPO1-dependent with LRPPRC interacting with XPO1 to form an EIF4E-dependent mRNA export complex (By similarity). Alters the composition of the cytoplasmic face of the nuclear pore to promote RNA export by reducing RANBP2 expression, relocalizing nucleoporin NUP214 and increasing expression of RANBP1 and RNA export factors DDX19 and GLE1 (By similarity). Promotes the nuclear export of cyclin CCND1 mRNA (By similarity). Promotes the nuclear export of NOS2/iNOS mRNA (PubMed:23471078). Promotes the nuclear export of MDM2 mRNA (PubMed:22684010). Promotes the export of additional mRNAs, including others involved in the cell cycle (By similarity). In the nucleus, binds to capped splice factor-encoding mRNAs and stimulates their nuclear export to enhance splice factor production by increasing their cytoplasmic availability to the translation machinery (By similarity). May also regulate splicing through interaction with the spliceosome in an RNA and m7G cap-dependent manner (By similarity). Also binds to some pre-mRNAs and may play a role in their recruitment to the spliceosome (By similarity). Promotes steady-state capping of a subset of coding and non-coding RNAs by mediating nuclear export of capping machinery mRNAs including RNMT, RNGTT and RAMAC to enhance their translation (By similarity). Stimulates mRNA 3'-end processing by promoting the expression of several core cleavage complex factors required for mRNA cleavage and polyadenylation, and may also have a direct effect through its interaction with the CPSF3 cleavage enzyme (By similarity). Rescues cells from apoptosis by promoting activation of serine/threonine-protein kinase AKT1 through mRNA export of NBS1 which potentiates AKT1 phosphorylation and also through mRNA export of AKT1 effectors, allowing for increased production of these proteins (By similarity).
EIF4EL1, EIF4F, EIF4E, Eukaryotic translation initiation factor 4E, eIF-4E, eIF4E, eIF-4F 25 kDa subunit, mRNA cap-binding protein
Rabbit Recombinant Monoclonal eIF4E antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Rat, Human samples. Cited in 4 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
The eIF4E protein also known as the eukaryotic translation initiation factor 4E functions mainly in the initiation phase of mRNA translation. It plays a role by binding to the 7-methylguanylate cap structure at the 5’ end of mRNA facilitating ribosome assembly on mRNA. The molecular weight of eIF4E is approximately 25 kDa and it is ubiquitously expressed in various tissues across eukaryotic organisms. Additionally phosphorylation of eIF4E affects its activity and is an important post-translational modification.
EIF4E regulates gene expression at the translational level and is a part of the eIF4F complex which also includes eIF4A and eIF4G. This complex is essential for the recruitment of ribosomes to the mRNA impacting the rate of protein synthesis. Changes in eIF4E levels can alter the translational efficiency of specific mRNAs thereby influencing cellular growth and proliferation.
EIF4E directly involves in the mTOR signaling pathway which regulates cell growth proliferation and survival. Within this pathway eIF4E interacts with proteins such as 4EBP1 which binds to eIF4E and inhibits its function in the absence of phosphorylation by mTOR. This interaction illustrates the control mechanism of protein synthesis under different cellular conditions connecting it to various signaling cascades.
EIF4E has been linked to cancer and neurodegenerative conditions. In cancer overexpression of eIF4E leads to increased translation of oncogenes facilitating tumorigenesis. The mTOR pathway proteins like mTOR and 4EBP1 are often dysregulated in these cases. Conversely its role in neurological disorders involves the aberrant regulation of synaptic proteins like PSD95 impacting neuronal communication and function.
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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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab33768 in NIH/3T3 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-eIF4E antibody [Y449] (ab33768)
Predicted band size: 25 kDa
Observed band size: 28 kDa
Exposure time: 3min
Exposure time:
Lane 1: 20 seconds.
Lanes 2-5: 8 seconds.
All lanes: Western blot - Anti-eIF4E antibody [Y449] (ab33768) at 1/1000 dilution
Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 4: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 5: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 28 kDa
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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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.
Intracellular 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, Goat Anti-Rabbit IgG H&L (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) analysis of mouse testis tissue labelling eIF4E with ab33768 at a dilution of 1/500. Antigen retrieval was performed Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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 rat testis tissue labelling eIF4E with ab33768 at a dilution of 1/500. Antigen retrieval was performed Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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.
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