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Purified anti-eIF4E Antibody

Pricing & Availability
Clone
P85D11B4 (See other available formats)
Regulatory Status
RUO
Other Names
Eukaryotic translation initiation factor 4E, eIF-4E, eIF-4F 25 kD subunit, mRNA cap-binding protein
Isotype
Mouse IgG2a, κ
Ave. Rating
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Product Citations
publications
A_P85D11B4_PURE_eIF4E_1_WB_101917.png
Total cell lysate from NIH3T3 cells (lane 1, 15 µg) and Jurkat cells (lane 2, 15 µg) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with purified anti-eIF4E antibody (clone P85D11B4, 0.5 µg /mL). Proteins were visualized using an HRP goat anti-mouse IgG antibody and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody (clone 2F1-1) was used as a loading control.
  • A_P85D11B4_PURE_eIF4E_1_WB_101917.png
    Total cell lysate from NIH3T3 cells (lane 1, 15 µg) and Jurkat cells (lane 2, 15 µg) were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with purified anti-eIF4E antibody (clone P85D11B4, 0.5 µg /mL). Proteins were visualized using an HRP goat anti-mouse IgG antibody and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody (clone 2F1-1) was used as a loading control.
  • B_P85D11B4_PURE_eIF4E_2_WB_070618
    Total cell lysates (15 µg protein) from K562 (lane 1), Molt4 (lane 2), HaCaT (lane 3) and HeLa (lane 4) cells were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 0.5 µg/mL (1:1000 dilution) of purified anti-eIF4E antibody (clone P85D11B4) (upper blot). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for anti-eIF4E antibody or 1:5000 diluted Direct-Blot HRP anti-β-Actin antibody, clone 2F1-1(lower blot). Lane M: Molecular weight ladder.
  • C_P85D11B4_PURE_eIF4E_2_IF_101917.png
    HeLa cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 0.5 µg/mL (1:1000) anti-eIF4E antibody (clone P85D11B4) overnight at 4°C followed by Alexa Fluor® 594 (red) conjugated goat anti-mouse IgG for one hour at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.
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693002 100 µg $265
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Description

The initiation step of protein translation requires a number of eukaryotic translation initiation factors (eIFs). Initially, the 40S ribosomal subunit associates with Met-tRNAi and a group of initiation factors, including eIF1, eIF1A, eIF2, eIF3, and eIF5 to form the 43S pre-initiation complex. This complex is then recruited to the 5'-end of capped mRNA by another complex of factors (known as eIF4F) consisting of eIF4E, eIF4G, and eIF4A. eIF4E is an indispensable element for cap-dependent translation initiation and plays a major role in recognition of the mRNA cap structure. It binds to eIF4G, which serves as a scaffolding protein to gather eIF4A (ATP-dependent RNA helicase), eIF4B, eIF3, and poly-A-binding protein to form the 48S complex. This complex then scans the mRNA for an initiation codon and, once this is located, the 60S ribosomal subunit joins to form the elongation-competent fully functional 80S. eIF4E is the least abundant of the cellular translation initiation components and it represents the rate-limiting element for the initiation process. Under resting conditions, eIF4E is kept in an inactive form bound to 4E-binding proteins (4E-BPs); once the latter have been phosphorylated through the PI3K-AKT-mTOR signaling pathway, eIF4E is released and binds eIF4G to launch translation.

  eIF4E is upregulated in various human tumors, and it contributes to transformation, tumorigenesis, and cancer progression. eIF4E overexpression facilitates translation of weak and highly structured mRNAs that typically encode proteins involved in cancer biology, such as proto-oncoproteins (e.g. cyclin D1, ornithine decarboxylase), angiogenesis factors (e.g. FGF-2, VEGF), and factors related to tumor invasiveness, such as MMP-9. Therefore, eIF4E serves as an attractive cancer drug target.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Purified recombinant full length of human eIF4E (AA: 1-217) expressed in E. coli.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/ml. For immunocytochemistry, a concentration range of 0.2 - 2.0 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone P85D11B4 does not cross-react with mouse (in-house tested).

Product Citations
  1. Wilde BR, et al. 2020. Cancer Metab. 8:27. PubMed
RRID
AB_2686932 (BioLegend Cat. No. 693002)

Antigen Details

Structure
217 amino acids long, and the predicted molecular weight is 25 kD.
Distribution

Cytoplasm

Function
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.
Interaction
eIF4E is a multi-subunit complex. It is composed of EIF4A, EIF4E and EIF4G1/EIF4G3. EIF4E is also known to interact with other partners, eg.EIF4ENIF1, EIF4A1, EIF4A2, NGDN, PIWIL2, CYFIP, FMR1, CLOCK, Lassa virus Z protein, MKNK2, LIMD1, WTIP, AJUBA, APOB
Biology Area
Cell Biology, Cell Cycle/DNA Replication, Protein Synthesis
Antigen References

1. Kapp LD, Lorsch JR. 2004. Annu. Rev. Biochem. 73: 657.
2. von der Haar T, et al. 2004. Nat. Struct. Mol. Biol. 11: 503.
3. Gingras AC, et al. 1999. Annu. Rev. Biochem. 68: 913.
4. De Benedetti A, Graff JR. 2004. Oncogene 23: 3189.
5. Zimmer SG, et al. 2000. Anticancer Res. 20:1343.
6. Jia Y, et al. 2012. Med. Res. Rev. 32: 786.
7. Graff JR, et al. 2007. J. Clin. Invest. 117: 2638.

Gene ID
1977 View all products for this Gene ID
UniProt
View information about eIF4E on UniProt.org

Related FAQs

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Go To Top Version: 2    Revision Date: 10/29/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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