Recombinant 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
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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.
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Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB HumanRecombinant fragment -
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.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
EP526Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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IHC-P |
Human
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WB |
Human
Recombinant fragment
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All applications |
Mouse
Rat
Drosophila melanogaster
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Application | Abreviews | Notes |
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WB | (1) |
1/5000 - 1/10000. Predicted molecular weight: 18 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt |
1/50 - 1/80.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/500.
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Notes |
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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
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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
modificationseIF-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
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Database links
- Entrez Gene: 1984 Human
- Entrez Gene: 276770 Mouse
- Entrez Gene: 287444 Rat
- Omim: 600187 Human
- SwissProt: P63241 Human
- SwissProt: P63242 Mouse
- SwissProt: Q3T1J1 Rat
- Unigene: 534314 Human
see all -
Alternative names
- eIF 4D antibody
- eIF 5A 1 antibody
- EIF 5A antibody
see all
Images
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Anti-eIF5A antibody [EP526Y] (ab32443) at 1/10000 dilution + Jurkat cell lysate.
Predicted band size: 18 kDa
Observed band size: 18 kDa -
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
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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.
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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 secondsBlocking buffer: 5% NFDM/TBST
This antibody shows low affinity in recognizing eIF5A2 recombinant protein.
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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.
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.
Datasheets and documents
References (12)
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