Anti-eIF4E (phospho S209) antibody
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(1 Publication)
Rabbit Polyclonal eIF4E phospho S209 antibody. Suitable for WB, ICC/IF and reacts with Rat, Human samples. Cited in 1 publication. Immunogen corresponding to Synthetic Peptide within Human EIF4E phospho S209.
View Alternative Names
EIF4EL1, EIF4F, EIF4E, Eukaryotic translation initiation factor 4E, eIF-4E, eIF4E, eIF-4F 25 kDa subunit, mRNA cap-binding protein
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-eIF4E (phospho S209) antibody (AB4774)
Immunofluorescence analysis using 70% confluent log phase U-87 MG cells labeled for Phospho-eIF4E pSer209 using ab4774. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ab4774 at 1 : 250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1 : 2000 for 45 minutes at room temperature (Panel A : green). Nuclei (Panel B : blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel C : red) was stained with Rhodamine Phalloidin (1 : 300). Panel D is a merged image showing cytoplasmic localization. Panel E is a no primary antibody control. The images were captured at 60X magnification.
- WB
Supplier Data
Western blot - Anti-eIF4E (phospho S209) antibody (AB4774)
Western blot analysis was performed on HeLa, HEK-293 (human cell lines) and L6 (rat cell line) cell lysates with various treatments, blotted for eIF4E (pS209) using ab4774. Two bands ~ 25 and 28 kDa band corresponding to eIF4E (pS209) was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate
All lanes:
Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
Lane 1:
HeLa whole cell extracts at 20 µg
Lane 2:
Serum starved HeLa at 20 µg
Lane 3:
HeLa Serum Starved for overnight followed by Serum Released at 20 µg
Lane 4:
HEK-293 at 20 µg
Lane 5:
HEK-293 treated for 30 minutes with 25 µg/mL of Anisomycin at 20 µg
Lane 6:
L6 at 20 µg
Lane 7:
L6 treated for 10 minutes with 200 ng/mL of Insulin at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 0.4 µg/mL
Predicted band size: 25 kDa
false
- WB
Unknown
Western blot - Anti-eIF4E (phospho S209) antibody (AB4774)
Extracts of HeLa cells were resolved by SDS-PAGE on a 4-20% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, either left untreated (1-4) or treated with lambda (?) phosphatase (5), and then incubated with the eIF4E (phospho S209) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with : no peptide (1,5), the nonphosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphoserine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Pierce SuperSignal™ method. The data show that only the peptide corresponding to eIF4E (phospho S209) blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody i
All lanes:
Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
Predicted band size: 25 kDa
false
Reactivity data
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Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
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Target data
Publications (1)
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Molecular cell 25:113-26 PubMed17218275
2007
Applications
Unspecified application
Species
Unspecified reactive species
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