Phospho-eIF4E (S209) Recombinant Rabbit Monoclonal Antibody [SU0396]
cat.: ET1608-66
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IP
Clonality: Monoclonal
Clone number: SU0396
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 25 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser209 of human eIF4E.
Positive control: Mouse spleen tissue lysate, rat spleen tissue lysate, NIH3T3 cell lysate, N2A, SH-SY5Y, 293, human pancreas tissue, rat spleen tissue, mouse brain tissue, mouse hippocampus tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IP

1:500-1:2,000
1:50-1:200
1:50-1:200
Use at an assay dependent concentration.
Uniprot #: SwissProt: P06730 Human | P63073 Mouse | P63074 Rat
Alternative names: AUTS19 CBP eIF 4E eIF 4F 25 kDa subunit EIF 4F eIF-4E eIF-4F 25 kDa subunit eIF4E EIF4E1 EIF4EL1 EIF4F Eukaryotic translation initiation factor 4 E Eukaryotic translation initiation factor 4E Eukaryotic translation initiation factor 4E like 1 IF4E_HUMAN Messanger RNA Cap Binding Protein eIF 4E MGC111573 mRNA cap binding protein mRNA cap-binding protein
Images
ET1608-66_1.jpg Fig1: Western blot analysis of Phospho-eIF4E (S209) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: mouse spleen tissue lysate
Lane 2: rat spleen tissue lysate
ET1608-66_2.jpg Fig2: Western blot analysis of Phospho-eIF4E (S209) on NIH-3T3 cell lysates.

Lane 1: NIH-3T3 cells, whole cell lysate, 10 μg /lane.
Lane 2: NIH-3T3 cells were treated with 2.8μg/ul lambda-PP for 30 minutes, whole cell lysates, 10 μg/lane.

Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody Anti-Phospho-eIF4E (S209) (ET1608-66, 1/500) and Anti-GAPDH antibody (ET1601-4, 1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Predicted band size: 25 kDa
Observed band size: 25 kDa
Exposure time: 2 minutes 29 seconds
ET1608-66_3.jpg Fig3: Western blot analysis of Phospho-eIF4E (S209) on rat spleen tissues lysates.

Lane 1: Rat spleen tissues, whole tissue lysates, 20 μg /lane.
Lane 2: Rat spleen tissues were treated with 2.8μg/ul lambda-PP for 30 minutes, whole tissue lysates, 20 μg/lane.

Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody Anti-Phospho-eIF4E (S209) (ET1608-66, 1/500) and Anti-GAPDH antibody (ET1601-4, 1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Predicted band size: 25 kDa
Observed band size: 25 kDa
Exposure time: 3 seconds
ET1608-66_4.jpg Fig4: ICC staining of Phospho-eIF4E (S209) in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-66_5.jpg Fig5: ICC staining of Phospho-eIF4E (S209) in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-66_6.jpg Fig6: ICC staining of Phospho-eIF4E (S209) in 293 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-66_7.jpg Fig7: 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.
ET1608-66_8.jpg Fig8: 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.
ET1608-66_9.jpg Fig9: 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.
ET1608-66_10.jpg Fig10: 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.
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