Invitrogen

Phospho-eIF4E (Ser209) Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

1 Published Figure

4 References

View all (41) eIF4E antibodies

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Phospho-eIF4E (Ser209) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Phospho-eIF4E pSer209 was done on 70% confluent log phase U-87 MG cells. 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 Phospho eIF4E pSer209 Rabbit Polyclonal Antibody (Product # 44-528G) 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 (Product # A27034) 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 (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 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.

Phospho-eIF4E (Ser209) Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (20 µg lysate) of HeLa (lane 1), Serum Starved HeLa (lane 2), HeLa Serum Starved for overnight followed by Serum Released (lane 3), HEK-293 (lane 4), HEK-293 treated for 30 minutes with 25 µg/mL of Anisomycin (lane 5), L6 (lane 6) and L6 treated for 10 minutes with 200 ng/mL of Insulin (lane 7). The blots were probed with Anti-eIF4E (pS209) Rabbit Polyclonal Antibody (Product # 44-528G, 1:500-1:2000 dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). 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 (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). 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 (Product # 32106).

Phospho-eIF4E (Ser209) Antibody in Immunoprecipitation (IP) — Immunoprecipitation of EIF4E pSer209 was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of EIF4E pSer209 polyclonal antibody (Product # 44-528G) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti- rabbit coated Dynabeads (Product # 11204D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a EIF4E pSer209 polyclonal antibody (Product # 44-528G) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

Phospho-eIF4E (Ser209) Antibody in RNA Immunoprecipitation (RIP) — RNA immunoprecipitation (RIP) western of EIF4E pSer209 was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of EIF4E pSer209 polyclonal antibody (Product # 44-528G) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti- rabbit coated Dynabeads (Product # 11204D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a EIF4E pSer209 polyclonal antibody (Product # 44-528G) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).

Phospho-eIF4E (Ser209) Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot of Phospho-eIF4E (Ser209) using Anti-eIF4E [pS209] Rabbit Polyclonal Antibody (Product # 44-528G), shows increase in expression of Phospho-eIF4E (Ser209) in HeLa cells Serum Starved for overnight followed by Serum Release,HEK-293 cells treated with Anisomycin and L6 cells treated with Insulin. {TM}

Product Details

44-528G

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 4 publications 4 publications
Immunocytochemistry (ICC/IF)	1:250	-
Immunoprecipitation (IP)	Assay-dependent	-
RNA Immunoprecipitation (RIP)	Assay-dependent	-

Product Specifications
Species Reactivity	Human, Mouse, Rabbit, Rat
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human eIF4E that contains serine 209. The sequence is conserved in mouse, rat and rabbit. View immunogen
Conjugate	Unconjugated
Form	Liquid
Purification	Antigen affinity chromatography
Storage buffer	Dulbecco's PBS, pH 7.3, with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20°C
RRID	AB_2533674

Target Information

eIF4E is involved in several cellular processes including enhanced translational efficiency, splicing, mRNA stability, and RNA nuclear export. All eukaryotic cellular mRNAs are blocked at their 5-prime ends with the 7-methylguanosine cap structure, m7GpppX (where X is any nucleotide). EIF4E is a eukaryotic translation initiation factor involved in directing ribosomes to the cap structure of mRNAs. It is a 24-kD polypeptide that exists as both a free form and as part of a multiprotein complex termed EIF4F. The EIF4E polypeptide is the rate-limiting component of the eukaryotic translation apparatus and is involved in the mRNA-ribosome binding step of eukaryotic protein synthesis. The other subunits of EIF4F are a 50-kD polypeptide, termed EIF4A, that possesses ATPase and RNA helicase activities, and a 220-kD polypeptide, EIF4G.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Identification of DNA response elements regulating expression of CCAAT/enhancer-binding protein (C/EBP) β and δ and MAP kinase-interacting kinases during early adipogenesis.

Adipocyte

Merrett JE,Bo T,Psaltis PJ,Proud CG

Published figure using Phospho-eIF4E (Ser209) polyclonal antibody (Product # 44-528G) in Western Blot

Tue Dec 01 00:00:00 EST 2020

Sinoporphyrin sodium is a promising sensitizer for photodynamic and sonodynamic therapy in glioma.

Oncology reports

An YW,Liu HQ,Zhou ZQ,Wang JC,Jiang GY,Li ZW,Wang F,Jin HT

44-528G was used in Western Blotting to explore the antitumor effects of sinoporphyrin sodium (DVDMS)‚mediated photodynamic therapy (PDT) and sonodynamic therapy (SDT) in glioma, and to reveal the underlying mechanisms.

Thu Oct 01 00:00:00 EDT 2020

Cotargeting MNK and MEK kinases induces the regression of NF1-mutant cancers.

The Journal of clinical investigation

Lock R,Ingraham R,Maertens O,Miller AL,Weledji N,Legius E,Konicek BM,Yan SC,Graff JR,Cichowski K

44-528G was used in western blot to analyze induction of the regression of NF1-mutant cancers by cotargeting MNK and MEK kinases

Wed Jun 01 00:00:00 EDT 2016

Autocrine Semaphorin3A stimulates eukaryotic initiation factor 4E-dependent RhoA translation in breast tumor cells.

Experimental cell research

Pan H,Bachelder RE

Fri Oct 15 00:00:00 EDT 2010

Bioinformatics

Protein Aliases: eIF-4E; eIF-4F 25 kDa subunit; Eukaryotic translation initiation factor 4E; eukaryotic translation initiation factor 4E-like 1; MGC111573; mRNA cap-binding protein; OTTHUMP00000219703; OTTHUMP00000219704; OTTHUMP00000219705

Gene Aliases: AUTS19; CBP; EG668879; EIF-4E; EIF4E; Eif4e-ps; EIF4E1; EIF4EL1; EIF4F; If4e

UniProt ID: (Human) P06730, (Rabbit) P29338, (Mouse) P63073, (Rat) P63074

Entrez Gene ID: (Human) 1977, (Rabbit) 100009338, (Mouse) 13684, (Rat) 117045

Molecular Function: translation factor translation initiation factor translational protein