eIF4E Antibody (YA463)

eIF4E Antibody (YA463) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to eIF4E.

IF4E proteins play multifaceted roles in cellular processes, coordinating important functions in the cytoplasm and nucleus. In the cytoplasm, it acts as a key initiator and regulator of protein synthesis. Furthermore, within the nucleus, IF4E is essential for exporting a portion of mRNA to the cytoplasm, facilitating processes such as RNA capping, processing, and splicing. As a component of the eIF4F protein complex, IF4E plays a critical role in recognizing the mRNA cap, unwinding 5' terminal secondary structure, and recruiting the mRNA to the ribosome. It binds to the 7-methylguanosine (m7G)-containing mRNA cap, thereby initiating protein synthesis and inducing mRNA secondary structure unwinding, thereby promoting ribosome binding. IF4E cooperates with EIF4G1 to counteract EIF1-EIF4G1-promoted scanning, which is critical for TISU translation, where TISU element recognition eliminates the need for scanning. In addition to translation initiation, IF4E also acts as a cytoplasmic regulator of translation and stability. It is part of the CYFIP1-EIF4E-FMR1 complex and mediates translation repression by binding to the mRNA cap. Furthermore, in P bodies, IF4E participates in a complex that stores translationally inactive mRNA and prevents its degradation. Its diverse functions extend to roles in spermatogenesis, neurogenesis, and nuclear-cytoplasmic transport of mRNA. Notably, the involvement of IF4E in mRNA export relies on its ability to bind the m7G cap and the presence of the EIF4E-sensitive element (4ESE). LRPPRC promotes interaction with 4ESE to form an EIF4E-dependent mRNA export complex. IF4E-dependent mRNA export, independent of ongoing protein or RNA synthesis, is XPO1-dependent and LRPPRC interacts with XPO1. The dynamic action of IF4E alters the composition of nuclear pores and promotes RNA export by regulating the expression of key factors. It significantly promotes the nuclear export of specific mRNAs, including the cell cyclins CCND1, NOS2/iNOS, and MDM2.

Free

P06730

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

Protein A affinity purified.

Cytoplasm, P-body, Stress granule, Nucleus.

Non-conjugated

Unmodified

AB_3102828

Epigenetics and Nuclear Signaling

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500; IHC-P: 1:200; ICC: 1:50; FC: 1:500-1:1000

• 
  Western blot analysis of extracts from NIH/3T3 (lane 2(20μg) , NIH/3T3 (lane 3(40μg),using eIF4E Antibody (HY-P80118). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of NIH3T3 cells labeling eIF4E with eIF4E Antibody (HY-P80118) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with eIF4E Antibody (HY-P80118) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 594-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8003, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of NIH3T3 cells labeling eIF4E with eIF4E Antibody (HY-P80118) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with eIF4E Antibody (HY-P80118)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 594-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8003,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using eIF4E Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80118, 1/200) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse brain tissue using eIF4E Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80118, 1/200) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-P, ICC/IF, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Recombinant protein within Human eIF4E aa 118-217/217.