Rabbit Anti-phospho-eIF4E (Ser209) antibody

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eIF4E(phospho S209); eIF4E(phospho Ser209); p-eIF4E(Ser209); EIF4F; eIF-4E; eIF 4E; CBP; eIF 4F 25 kDa subunit; CBP; eIF 4E; eIF 4F 25 kDa subunit; EIF 4F; EIF4E1; EIF4EL1; EIF4F; Eukaryotic Translation Initiation Factor 4 E; Eukaryotic translation initia

Cat:

SL2446R

Species Reactivity：

Human,Mouse,Rat,Zebrafish,(predicted: Chicken,Dog,Pig,Cow,Horse,Rabbit,)

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human eIF4E around the phosphorylation site of Ser209:SG(p-S)TT

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=0.2ug/testIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: NIH/3T3(Mouse) Cell Lysate at 30 ugPrimary: Anti-phospho-eIF4E (Ser209) (SL2446R) at 1/500 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 24 kDObserved band size: 28 kD Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (SL2446R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (SL2446R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (SL2446R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Blank control:Molt-4. Primary Antibody (green line): Rabbit Anti-phospho-eIF4E (Ser209) antibody (SL2446R) Dilution: 0.2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 0.2μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 20% PBST for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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eIF4E, a protein modulates translation of maternal mRNAs in early embryos before the onset of zygotic transcription. eIF4E also influences the overall rate of translation. eIF4E binds to the 7 methyl GTP cap structure of eukaryotic mRNAs. Phosphorylation of eIF4E on serine 209 regulates the affinity of this protein for the 7 methyl GTP cap and/or RNA. Phosphorylation also enhances the interaction of eIF4E with eIF4G, which form a complex known as eIF4F. eIF4E phosphorylation is correlated with increased translational rate in a number of cell types. Several kinases are currently being investigated as potential regulators of eIF4E including PKC and/or the MAP kinase activated Mnk.

Function:
Its translation stimulation activity is repressed by binding to the complex CYFIP1-FMR1 (By similarity). 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. Component of the CYFIP1-EIF4E-FMR1 complex which binds to the mRNA cap and mediates translational repression. In the CYFIP1-EIF4E-FMR1 complex this subunit mediates the binding to the mRNA cap.

Subunit:
eIF4F is a multi-subunit complex, the composition of which varies with external and internal environmental conditions. It is composed of at least EIF4A, EIF4E and EIF4G1/EIF4G3. EIF4E is also known to interact with other partners. The interaction with EIF4ENIF1 mediates the import into the nucleus. Nonphosphorylated EIF4EBP1, EIF4EBP2 and EIF4EBP3 compete with EIF4G1/EIF4G3 to interact with EIF4E; insulin stimulated MAP-kinase (MAPK1 and MAPK3) phosphorylation of EIF4EBP1 causes dissociation of the complex allowing EIF4G1/EIF4G3 to bind and consequent initiation of translation. Rapamycin can attenuate insulin stimulation, mediated by FKBPs. Interacts mutually exclusive with EIF4A1 or EIF4A2. Interacts with NGDN and PIWIL2. Component of the CYFIP1-EIF4E-FMR1 complex composed of CYFIP, EIF4E and FMR1. Interacts directly with CYFIP1 (By similarity). Interacts with Lassa virus Z protein. Binds to MKNK2 in nucleus. Interacts with LIMD1, WTIP and AJUBA.

Subcellular Location:
Cytoplasm, P-body.

Post-translational modifications:
Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.

Similarity:
Belongs to the eukaryotic initiation factor 4E family.

SWISS:
P06730

Gene ID:
1977

Database links:

Entrez Gene: 1977 Human

Entrez Gene: 13684 Mouse

Entrez Gene: 117045 Rat

Omim: 13388 Human

SwissProt: P06730 Human

SwissProt: P63073 Mouse

SwissProt: P63074 Rat

Unigene: 249718 Human

Unigene: 3941 Mouse

Unigene: 11275 Rat

EIF4E是蛋白合成的一个主要调控因子，它表达量增加会促进多种肿瘤的生长的增值、分化，因此EIF4E是一种非常重要的抗癌药物靶标。 Picture

Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (SL2446R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (SL2446R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-eIF4E (Ser209)) Polyclonal Antibody, Unconjugated (SL2446R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control:Molt-4.
Primary Antibody (green line): Rabbit Anti-phospho-eIF4E (Ser209) antibody (SL2446R)
Dilution: 0.2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 0.2μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 20% PBST for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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