Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

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Overview

  • Product name

    Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free
    See all eIF4EBP1 primary antibodies

  • Description

    Rabbit monoclonal [Y329] to eIF4EBP1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, IHC-P, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human eIF4EBP1 aa 1-100 (N terminal). The exact sequence is proprietary.

  • General notes

    Ab173370 is the carrier-free version of ab32024. 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.

    ab173370 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 ab173370 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
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 13 kDa).

Target

  • Function

    Regulates eIF4E activity by preventing its assembly into the eIF4F complex. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways.

  • Sequence similarities

    Belongs to the eIF4E-binding protein family.

  • Post-translational
    modifications

    Phosphorylated on serine and threonine residues in response to insulin, EGF and PDGF. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70 is regulated by mTORC1. Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 4E-BP1 antibody
    • 4EBP1 antibody
    • 4EBP1_HUMAN antibody
    • BP 1 antibody
    • eIF4E binding protein 1 antibody
    • eIF4E-binding protein 1 antibody
    • Eif4ebp1 antibody
    • Eukaryotic translation initiation factor 4E-binding protein 1 antibody
    • PHAS-I antibody
    • PHASI antibody
    • Phosphorylated heat- and acid-stable protein regulated by insulin 1 antibody
    see all

Images

  • Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-EIF4EBP1   knockout cells (red line) stained with ab32024. The cells were fixed with 80% methanol (5 min)  and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab32024, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-EIF4EBP1  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    This antibody can also be used in HAP1 cells fixed with 4% formaldehyde (10 min) , permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32024).

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling eIF4EBP1 with Purified ab32024 at 1/500 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32024).

  • Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Overlay histogram showing HT1080 cells stained with ab32024 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32024, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HT1080 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32024).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

    ab32024, at a 1/100 dilution, staining human eIF4EBP1 in colon carcinoma by Immunohistochemistry, Paraffin embedded tissue

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32024).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

References

ab173370 has not yet been referenced specifically in any publications.

Publishing research using ab173370? Please let us know so that we can cite the reference in this datasheet.

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