Recombinant
RabMAb

Recombinant Anti-eIF4E (phospho S209) antibody [EP2151Y] - BSA and Azide free (ab183301)

Overview

  • Product name

    Anti-eIF4E (phospho S209) antibody [EP2151Y] - BSA and Azide free
    See all eIF4E primary antibodies
  • Description

    Rabbit monoclonal [EP2151Y] to eIF4E (phospho S209) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Dot blot, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic peptide within Human eIF4E (phospho S209). The exact sequence is proprietary.

  • Positive control

    • 293 cell lysates, untreated or treated with AP; human breast carcinoma tissue.
  • General notes

    Ab183301 is the carrier-free version of ab76256. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab183301 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab183301 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Predicted molecular weight: 25 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Dot blot Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • 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.
  • Sequence similarities

    Belongs to the eukaryotic initiation factor 4E family.
  • Post-translational
    modifications

    Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
  • Information by UniProt
  • Database links

  • Alternative names

    • 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
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of untreated, 20% serum treated and 20% serum + LP treated NIH/3T3 cells labelling eIF4E (phospho S209) with ab76256 (left) and eIF4E with ab33766 (right) both at a dilution of 1/500.

    Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    The image shows increased cytoplasmic staining after 20% serum treatment on NIH3T3 cells when compared with no serum treated cells. The LP treatment decreased the increased cytoplasmic staining caused by 20% serum.

    ab33766 was used as a Pan control for ab76256. The results showed cytoplasmic staining on no serum, 20% serum and 20% serum +LP treated NIH3T3 cells.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76256).

  • ab76256 (purified) at 1/40 immunoprecipitating eIF4E (phospho S209) in HEK293 whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1,500) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76256).

  • Dot blot analysis of eIF4E (pS209) peptide (Lane 1) and eIF4E non-phospho peptide (Lane 2) labelling eIF4E (pS209) with purified ab76256 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76256).

  • Immunocytochemistry/Immunofluorescence analysis of serum starved HEK293 cells treated with CGP 57380 ab120365) labelling eIF4E (phospho S209) with unpurified ab32124 at 1/100. Decrease in eIF4E (phospho S209) expression correlates with increased concentration of CGP 57380, as described in literature.
    The cells were incubated at 37°C for 1h in media containing different concentrations of ab120365 (CGP 57380) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with unpurified ab76256 was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76256).

  • This ICC/IF data was generated using the same anti-eIF4E (phospho S209) antibody clone, EP2151Y, in a different buffer formulation (cat# ab76256).

    Immunocytochemistry/Immunofluorescence analysis of untreated, 20% serum treated and 20% serum + LP treated NIH/3T3 cells labelling eIF4E (phospho S209) with ab76256 (left) and eIF4E with ab33766 (right) both at a dilution of 1/500.

    Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    The image shows increased cytoplasmic staining after 20% serum treatment on NIH3T3 cells when compared with no serum treated cells. The LP treatment decreased the increased cytoplasmic staining caused by 20% serum.

    ab33766 was used as a Pan control for ab76256. The results showed cytoplasmic staining on no serum, 20% serum and 20% serum +LP treated NIH3T3 cells.

  • This IHC data was generated using the same anti-eIF4E (phospho S209) antibody clone, EP2151Y, in a different buffer formulation (cat# ab76256).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling eIF4E with purified ab76256 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

References

This product has been referenced in:

  • Chung CH  et al. Butein Inhibits Angiogenesis of Human Endothelial Progenitor Cells via the Translation Dependent Signaling Pathway. Evid Based Complement Alternat Med 2013:943187 (2013). WB ; Rat . Read more (PubMed: 23840271) »
  • Martineau Y  et al. Pancreatic tumours escape from translational control through 4E-BP1 loss. Oncogene N/A:N/A (2013). Read more (PubMed: 23563181) »
See all 6 Publications for this product

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