Anti-EphA3 (extracellular) Antibody

Ephrin type-A receptor 3, EPH-like kinase 4, EK4, Tyrosine-protein kinase TYRO4
    Cat #: AER-013
    Alternative Name Ephrin type-A receptor 3, EPH-like kinase 4, EK4, Tyrosine-protein kinase TYRO4
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    • Peptide (C)RARGTNVTISSLK, corresponding to amino acid residues 488-500 of human EphA3 (Accession P29320). Extracellular, N-terminus.
    • Anti-EphA3 (extracellular) Antibody
    Accession (Uniprot) Number P29320
    Gene ID 2042
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse – identical.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype Rabbit IgG.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Negative control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for two weeks. For longer periods, it should be stored at -20°C.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: if, ifc, ih, wb
    May also work in: ic*, ip*, lci*
    Western blot
    • Anti-EphA3 (extracellular) Antibody
      Western blot analysis of human Jurkat T-cell leukemia cell line lysate (lanes 1 and 5), human K562 chronic myelogenous leukemia cell line lysate (lanes 2 and 6), human Malme-3M melanoma cell line lysate (lanes 3 and 7) and human HT-29 colorectal adenocarcinoma cell line lysate (lanes 4 and 8):
      1-4. Anti-EphA3 (extracellular) Antibody (#AER-013), (1:200).
      5-8. Anti-EphA3 (extracellular) Antibody, preincubated with the negative control antigen.
    • Anti-EphA3 (extracellular) Antibody
      Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain membranes:
      1,2. Anti-EphA3 (extracellular) Antibody (#AER-013), (1:200).
      3,4. Anti-EphA3 (extracellular) Antibody, preincubated with the negative control antigen.
    • Anti-EphA3 (extracellular) Antibody
      Expression of EphA3 in rat and mouse hippocampus
      Immunohistochemical staining of perfusion-fixed frozen rat and mouse brain sections with Anti-EphA3 (extracellular) Antibody (#AER-013), (1:200), followed by goat anti-rabbit-AlexaFluor-488. A. EphA3 staining (green) in rat hippocampal CA1 region is detected in the pyramidal layer (P). B. In mouse hippocampal CA1 region, EphA3 immunoreactivity (green) is observed in the pyramidal layer (P). Cell nuclei are stained with DAPI (blue).
    Indirect flow cytometry
    • Anti-EphA3 (extracellular) Antibody
      Cell surface detection of EphA3 in live intact human Jurkat T-cell leukemia cells:
      ___ Cells.
      ___ Cells + goat-anti-rabbit-FITC.
      ___ Cells + Anti-EphA3 (extracellular) Antibody (#AER-013), (5µg) + goat-anti-rabbit-FITC.
    1. Offenhäuser, C. et al. (2018) Cancers (Basel) 10, 12.
    2. Forse, G.J. et al. (2015) PLoS One 10, e0127081.
    3. Nasri, B. et al. (2017) BMC Clin. Pathol. 17, 8.
    4. Andretta, E. et al. (2017) Sci. Rep. 7, 41576.
    Scientific background

    Eph receptors are the largest family of receptor tyrosine kinases (RTKs). Based on sequence similarity and the type of ephrin ligands that they bind, they are classified into A and B sub-classes. EphA receptors bind ephrin-A ligand while EphB bind ephrin-B ligand.

    Eph receptors play an important role during embryonic development and are involved in several processes including cell adhesion, cell migration, and tissue boundary formation. Eph receptors are typically expressed at low levels in normal adult tissues1,2.

    EphA3 (Ephrin type-A receptor 3) structure includes a ligand-binding domain (LBD) at the N-terminus followed by a cysteine-rich region and two fibronectin III repeats. The receptor-binding domain of the ephrins is connected through a short linker segment to a GPI-anchor or a transmembrane segment3.

    EphA3 was first identified as a surface antigen on pre-B lymphoblastic leukemia cells. High EphA3 levels have been detected in a number of hematological cancers and solid tumors such as lung cancers, melanomas, gastric carcinoma, leukemia, in B and T cell malignancies and in glioblastoma multiforme. In addition, high levels of EphA3 are implicated in lymph node metastasis, advanced stages of colorectal cancer, and prostate cancer1,3,4.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 03/02/2020

    Anti-EphA3 (extracellular) Antibody (#AER-013) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunohistochemistry and live cell flow cytometry applications. It has been designed to recognize EphA3 from human, rat, and mouse samples.

    For research purposes only, not for human use
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