PRODUCT CODE: ET1608-66

Recombinant Phospho eIF4E (S209) Monoclonal Antibody (ET1608-66)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

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.<br />
 Positive control: <br />
 Lane 1: mouse spleen tissue lysate<br />
 Lane 2: rat spleen tissue lysate
  • 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.<br />
 Positive control: <br />
 Lane 1: mouse spleen tissue lysate<br />
 Lane 2: rat spleen tissue lysate
  • 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).
  • 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).
  • 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).
  • Immunohistochemical analysis of paraffin-embedded human 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.
  • 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.
  • Immunohistochemical analysis of paraffin-embedded mouse 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.
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

Applications

  • WB

  • ICC

  • IHC-P

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho eIF4E (S209) Monoclonal Antibody (ET1608-66)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser209 of human eif4e.

Host

Rabbit

Modification

Phospho

Modification Site

S209

Positive Control

Mouse spleen tissue lysate, rat spleen tissue lysate, N2A, SH-SY5Y, 293, human spleen tissue, rat spleen tissue, mouse spleen tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SU0396

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

25 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-eIF4E (S209)

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; EIF4F antibody; Eukaryotic translation initiation factor 4 E antibody; Eukaryotic translation initiation factor 4E antibody; Eukaryotic translation initiation factor 4E like 1 antibody; IF4E_HUMAN antibody; Messanger RNA Cap Binding Protein eIF 4E antibody; MGC111573 antibody; mRNA cap binding protein antibody; mRNA cap-binding protein antibody

SEQUENCE SIMILARITIES

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.

FUNCTION

The initiation of protein synthesis in eukaryotic cells is regulated by interactions between protein initiation factors and RNA molecules. The eukaryotic initiation complex eIF4F exists in vitro as a trimeric complex of eIF4E, eIF4A and eIF4G. Together, the complex allows ribosome binding to mRNA by inducing the unwinding of mRNA secondary structures. eIF4E binds to the mRNA "cap" during an early step in the initiation of protein synthesis. eIF4A acts as an ATP-dependent RNA helicase. eIF4G acts as a bridge between eIF4E, eIF4A and the eIF3 complex.