Phospho-eIF4E (S209) Recombinant Rabbit Monoclonal Antibody [SU0396] - BSA and Azide free
Catalog# HA750160
Phospho-eIF4E (S209) Recombinant Rabbit Monoclonal Antibody [SU0396] - BSA and Azide free
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WB
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IF-Cell
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Phospho-eIF4E (S209) Recombinant Rabbit Monoclonal Antibody [SU0396] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser209 of human eIF4E.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, IP, FC
Target Molecular Weight
Predicted band size: 25 kDa
Positive Control
HeLa cell lysate, HeLa treated with 50nM dexamethasone for 16 hours cell lysate, NIH/3T3 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, NIH/3T3, human pancreas tissue, mouse brain tissue, mouse hippocampus tissue, rat spleen tissue, 293T.
Conjugation
unconjugated
Clone Number
SU0396
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
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.In addition to its role in translation initiation, also acts as a regulator of translation and stability in the cytoplasm.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.Component of a multiprotein complex that sequesters and represses translation of proneurogenic factors during neurogenesis.In P-bodies, component of a complex that mediates the storage of translationally inactive mRNAs in the cytoplasm and prevents their degradation.May play an important role in spermatogenesis through translational regulation of stage-specific mRNAs during germ cell development.
Background References
1. Zhang P et al. p53, MDM2, eIF4E and EGFR expression in nasopharyngeal carcinoma and their correlation with clinicopathological characteristics and prognosis: A retrospective study. Oncol Lett 9:113-118 (2015).
2. Zheng J et al. Phosphorylated Mnk1 and eIF4E are associated with lymph node metastasis and poor prognosis of nasopharyngeal carcinoma. PLoS One 9:e89220 (2014).
Sequence Similarity
Belongs to the eukaryotic initiation factor 4E family.
Post-translational Modification
Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
Subcellular Location
Cytoplasm, Nucleus.
Synonyms
AUTS19 antibody
CBP antibody
eIF 4E antibody
eIF 4F 25 kDa subunit antibody
EIF 4F antibody
eIF-4E antibody
eIF-4F 25 kDa subunit antibody
eIF4E antibody
EIF4E1 antibody
EIF4EL1 antibody
ExpandImages
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☑ Cell treatment (CT)
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Western blot analysis of Phospho-eIF4E (S209) on different lysates with Rabbit anti-Phospho-eIF4E (S209) antibody (ET1608-66) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: Mouse spleen tissue lysate
Lane 5: Rat spleen tissue lysate
Lane 6: HeLa treated with 50nM dexamethasone for 16 hours cell lysate, then the membrane treated with λpp for 1 hour
Lysates/proteins at 20 µg/Lane.
Predicted band size: 25 kDa
Observed band size: 25 kDa
Exposure time: 42 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-66) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of NIH/3T3 cells labeling Phospho-eIF4E (S209) with Rabbit anti-Phospho-eIF4E (S209) antibody (ET1608-66) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-eIF4E (S209) antibody (ET1608-66) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Phospho-eIF4E (S209) antibody (ET1608-66) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-66) at 1/200 dilution for 1 hour at room temperature. 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. -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-eIF4E (S209) antibody (ET1608-66) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-66) at 1/200 dilution for 1 hour at room temperature. 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. -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-Phospho-eIF4E (S209) antibody (ET1608-66) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-66) at 1/200 dilution for 1 hour at room temperature. 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. -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat spleen tissue using anti-Phospho-eIF4E (S209) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-66, 1/50) for 30 minutes at room temperature. 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. -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Phospho-eIF4E (S209) was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with ET1608-66 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1608-66 at 1/2,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: NIH/3T3 cell lysate (input)
Lane 2: ET1608-66 IP in NIH/3T3 cell lysate
Lane 3: Rabbit IgG instead of ET1608-66 in NIH/3T3 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 15 seconds; ECL: K1801 -
This data was developed using ET1608-66, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 293T cells labeling Phospho-eIF4E (S209).
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-66, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Products with the same target and pathway
Phospho-eIF4E (S209) Recombinant Rabbit Monoclonal Antibody [SU0396]
Application: WB,IF-Cell,IHC-P,IP,FC
Reactivity: Human,Mouse,Rat
Conjugate: unconjugated
