Anti-EphA1 (extracellular) Antibody

Ephrin type-A receptor 1, Tyrosine-protein kinase receptor EPH, EPHA1
    Cat #: AER-011
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)KKEPRQLELTWAGSR, corresponding to amino acid residues 457-471 of human EphA1 (Accession P21709). Extracellular, N-terminus.
    Accession (Uniprot) Number P21709
    Gene ID 2041
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, rat – identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    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).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, lci, wb
    May also work in: ih, ip
    Western blot
    Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
    1, 2. Anti-EphA1 (extracellular) Antibody (#AER-011), (1:200).
    3, 4. Anti-EphA1 (extracellular) Antibody, preincubated with the control peptide antigen.
    Western blot analysis of human Jurkat T cell leukemia cells (lanes 1 and 5); human HeLa cervix adenocarcinoma cells (lanes 2 and 6); human SH-SY5Y neuroblastoma cells (lanes 3 and 7) and human MCF-7 breast adenocarcinoma cells (lanes 4 and 8):
    1- 4. Anti-EphA1 (extracellular) Antibody (#AER-011), (1:200).
    5- 8. Anti-EphA1 (extracellular) Antibody, preincubated with the control peptide antigen.
    Indirect flow cytometry
    Indirect flow cytometry analysis of live intact human Jurkat T cell leukemia cell line:
    ___ Unstained cells + goat-anti-rabbit-PE.
    ___ Cells + Anti-EphA1 (extracellular) Antibody (#AER-011), (1:20) + goat-anti-rabbit-PE.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of EphA1 in human MCF-7 cells
    Immunocytochemical staining of live intact MCF-7 breast adenocarcinoma cells. Cells were stained with Anti-EphA1 (extracellular) Antibody (#AER-011), (1:25) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live image of the cells stained in A. C. Merge image of A and B.
    References
    1. Chen, Y. et al. (2012) Cell. Signal. 24, 606.
    2. Pasquale, E.B. (2008) Cell 133, 38.
    3. Pasquale, E.B. (2005) Mol. Cell Biol. 6, 462.
    4. Du, J. et al. (2007) Curr. Pharm. Des. 13, 2507.
    5. Wu, J. and Luo, H. (2005) Curr. Opin. Hematol. 12, 292.
    6. Ireton, R.C. and Chen, J. (2005) Curr. Cancer Drug Targets 5, 149.
    7. Noren, N.K. and Pasquale, E.B. (2007) Cancer Res. 67, 3994.
    Scientific background

    Eph receptors are the largest family of receptor tyrosine kinases (RTKs). EphA receptors (EphA1-10) bind ephrinA ligands which are GPI-linked proteins and EphB receptors (EphB1-6) bind ephrinB ligands which are membrane protein with one transmembrane domain1,2. Within each subfamily, interactions between receptor and ligand are promiscuous. In addition, Eph receptors can also bind ephrins from the other class2. Forward and reverse signaling through Eph receptors is a unique characteristic to this RTK since ephrins are physically linked to the plasma membrane3.

    Structurally, Eph receptors contain an extracellular ligand-binding domain, a transmembrane domain and an intracellular C-terminal domain responsible for intracellular signaling1. Forward Eph receptor signaling involves autophosphorylation of the receptor via a tyrosine kinase domain, as well as phosphorylation of other proteins. Known effectors of the forward signaling include Src kinase and Ras/Rho GTPases2. Much less is known about the reverse signaling mediated by Eph receptors. Besides from acting independently, Eph receptors can also signal in concert with other receptors. For example, Eph receptors cooperate with FGF receptor, NMDA ligand-gated ion channel and chemokine G-protein coupled receptor2.

    Biological activities attributed to the Eph receptor-ephrin signaling module include establishing neuronal connections, mediating neuronal plasticity and repair following neuronal injury2,4. Eph receptors may also have a role in the immune system5.

    Eph receptors are expressed in the developing nervous system, and in the adult brain. It is also detected in the pancreas, intestine, bone and lymphocytes. In cancer cells, Eph receptors and ephrins are overexpressed2,6,7. They are also implicated in neurodegenerative disorders like Alzheimer’s disease1.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, 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: 06/12/2018

    Anti-EphA1 (extracellular) Antibody (#AER-011) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunocytochemistry, live cell imaging, and indirect live cell flow cytometry applications. It has been designed to recognize EphA1 from rat, mouse, and human samples.

    For research purposes only, not for human use