RecombinantRabMAb

Recombinant Anti-eIF5A antibody [EP526Y] (ab32443)

Reviews (1)Q&A (1)References (12)
Western blot - Anti-eIF5A antibody [EP526Y] (ab32443)
  • Immunocytochemistry/ Immunofluorescence - Anti-eIF5A antibody [EP526Y] (ab32443)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)
  • Western blot - Anti-eIF5A antibody [EP526Y] (ab32443)
  • Flow Cytometry - Anti-eIF5A antibody [EP526Y] (ab32443)
  • Anti-eIF5A antibody [EP526Y] (ab32443)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP526Y] to eIF5A
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Human, Recombinant fragment

Overview

  • Product name

    Anti-eIF5A antibody [EP526Y]
    See all eIF5A primary antibodies

  • Description

    Rabbit monoclonal [EP526Y] to eIF5A

  • Host species

    Rabbit

  • Specificity

    This antibody shows low affinity in recognizing eIF5A2 recombinant protein.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    Recombinant fragment
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human eIF5A aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P63241

  • Positive control

    • Jurkat and HeLa whole cell lysate (ab150035) and Adenocarcinoma of human uterus slides.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32443 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
Recombinant fragment
All applications
Mouse
Rat
Drosophila melanogaster
Application Abreviews Notes
WB (1)
1/5000 - 1/10000. Predicted molecular weight: 18 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
1/50 - 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/500.
Notes
WB
1/5000 - 1/10000. Predicted molecular weight: 18 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
1/50 - 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/500.

Target

  • Function

    mRNA-binding protein involved in translation elongation. Has an important function at the level of mRNA turnover, probably acting downstream of decapping. Involved in actin dynamics and cell cycle progression, mRNA decay and probably in a pathway involved in stress response and maintenance of cell wall integrity. With syntenin SDCBP, functions as a regulator of p53/TP53 and p53/TP53-dependent apoptosis. Regulates also TNF-alpha-mediated apoptosis. Mediates effects of polyamines on neuronal process extension and survival. May play an important role in brain development and function, and in skeletal muscle stem cell differentiation. Also described as a cellular cofactor of human T-cell leukemia virus type I (HTLV-1) Rex protein and of human immunodeficiency virus type 1 (HIV-1) Rev protein, essential for mRNA export of retroviral transcripts.

  • Tissue specificity

    Expressed in umbilical vein endothelial cells and several cancer cell lines (at protein level).

  • Sequence similarities

    Belongs to the eIF-5A family.

  • Post-translational
    modifications

    eIF-5A seems to be the only eukaryotic protein to have an hypusine residue which is a post-translational modification of a lysine by the addition of a butylamino group (from spermidine).

  • Cellular localization

    Cytoplasm. Nucleus. Endoplasmic reticulum membrane. Nucleus > nuclear pore complex. Hypusine modification promotes the nuclear export and cytoplasmic localization and there was a dynamic shift in the localization from predominantly cytoplasmic to primarily nuclear under apoptotic inducing conditions.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • eIF 4D antibody
    • eIF 5A 1 antibody
    • EIF 5A antibody
    • eIF 5A1 antibody
    • eIF-4D antibody
    • eIF-5A antibody
    • eIF-5A-1 antibody
    • eIF-5A1 antibody
    • eIF4D antibody
    • eif5a antibody
    • EIF5A1 antibody
    • eIF5AI antibody
    • Eukaryotic initiation factor 5A antibody
    • Eukaryotic initiation factor 5A isoform 1 antibody
    • Eukaryotic translation initiation factor 5A 1 antibody
    • Eukaryotic translation initiation factor 5A antibody
    • Eukaryotic translation initiation factor 5A-1 antibody
    • IF5A1_HUMAN antibody
    • MGC104255 antibody
    • MGC99547 antibody
    • Rev binding factor antibody
    • Rev-binding factor antibody
    • uORF A antibody
    • uORF antibody
    see all

Images

  • Western blot - Anti-eIF5A antibody [EP526Y] (ab32443)
    Western blot - Anti-eIF5A antibody [EP526Y] (ab32443)
    Anti-eIF5A antibody [EP526Y] (ab32443) at 1/10000 dilution + Jurkat cell lysate.

    Predicted band size: 18 kDa
    Observed band size: 18 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF5A antibody [EP526Y] (ab32443)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF5A antibody [EP526Y] (ab32443)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling eIF5A with purified ab32443 at 1/500. Cells were fixed with 4% Paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei couterstained with DAPI (blue). 

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)

    ab32443 at a 1:250 dilution staining eIF5A in human adenocarcinoma of uterus using Immunohistochemistry, Paraffin Embedded Tissue.

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

  • Western blot - Anti-eIF5A antibody [EP526Y] (ab32443)
    Western blot - Anti-eIF5A antibody [EP526Y] (ab32443)
    All lanes : Anti-eIF5A antibody [EP526Y] (ab32443) at 1/2000 dilution

    Lane 1 : Recombinant Human eIF5A protein (ab87457)
    Lane 2 : Recombinant Human eIF5A2 protein (ab99140)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 18 kDa
    Observed band size: 15,20 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 seconds


    Blocking buffer: 5% NFDM/TBST

    This antibody shows low affinity in recognizing eIF5A2 recombinant protein.

  • Flow Cytometry - Anti-eIF5A antibody [EP526Y] (ab32443)
    Flow Cytometry - Anti-eIF5A antibody [EP526Y] (ab32443)
    Overlay histogram showing Jurkat cells stained with ab32443 (red line). The cells were fixed with methanol (5 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 (ab32443, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Anti-eIF5A antibody [EP526Y] (ab32443)
    Anti-eIF5A antibody [EP526Y] (ab32443)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (12)

Publishing research using ab32443? Please let us know so that we can cite the reference in this datasheet.

ab32443 has been referenced in 12 publications.

  • Gao YF  et al. LncRNA FOXD1-AS1 acts as a potential oncogenic biomarker in glioma. CNS Neurosci Ther 26:66-75 (2020). PubMed: 31102349
  • Wang Z  et al. EIF5A regulates proliferation and chemoresistance in pancreatic cancer through the sHH signalling pathway. J Cell Mol Med 23:2678-2688 (2019). PubMed: 30761741
  • Fujimura K  et al. KRAS Oncoprotein Expression Is Regulated by a Self-Governing eIF5A-PEAK1 Feed-Forward Regulatory Loop. Cancer Res 78:1444-1456 (2018). PubMed: 29321164
  • Zhang J  et al. EIF5A1 promotes epithelial ovarian cancer proliferation and progression. Biomed Pharmacother 100:168-175 (2018). WB . PubMed: 29428664
  • Muñoz-Soriano V  et al. Evolutionary conserved role of eukaryotic translation factor eIF5A in the regulation of actin-nucleating formins. Sci Rep 7:9580 (2017). IHC ; Drosophila melanogaster . PubMed: 28852021
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

Will this cross-react with human EIF5A2?
http://www.uniprot.org/uniprot/Q9GZV4

Read More
Answer

Thank you for contacting us.

The immunogen shares 74% identity with human EIF5A2, so it is not predicted to cross-react.

Please do not hesitate to contact us if you need any more advice or information.

Read More

Western blot abreview for Anti-eIF5A antibody [EP526Y]

 Good
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116 colon cancer cell line)
Loading amount
30 µg
Specification
HCT116 colon cancer cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% in TBST
Read More

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Submitted Nov 07 2006

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