Human EphA2 Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human EphA2 in direct ELISAs and Western blots. In direct ELISAs, approximately 45% cross-reactivity with recombinant mouse (rm) EphA2 is observed and less than 1% cross-reactivity with recombinant human (rh) EphA1, rhEphA3, rhEphA4, rhEphA5, rhEphA6, rhEphA7 and rhEphA10 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human EphA2
    Gln25-Asn534
    Accession # P29317
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Knockout Validated
    1 µg/mL
    A431 human epithelial carcinoma parental cell line and EphA2 knock out A431 cell line
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human EphA2 by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma parental cell line and EphA2 knock out (KO) A431 cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human EphA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3035) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for EphA2 at approximately 110 kDa (as indicated), but not detectable in the knockout A431 cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of EphA2 in A431 Human Cell Line by Flow Cytometry. A431 human epithelial carcinoma cell line was stained with Goat Anti-Human EphA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3035, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.
Immunohistochemistry
EphA2 in Human Ovarian Cancer Tissue. EphA2 was detected in immersion fixed paraffin-embedded sections of human ovarian cancer tissue using Goat Anti-Human EphA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3035) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane of cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: EphA2

EphA2, also known as Eck, Myk2, and Sek2, is a member of the Eph receptor tyrosine kinase family which binds Ephrins A1, 2, 3, 4, and 5 (1-4). A and B class Eph proteins have a common structural organization. The human EphA2 cDNA encodes a 976 amino acid (aa) precursor including a 24 aa signal sequence, a 510 aa extracellular domain (ECD), a 24 aa transmembrane segment, and a 418 aa cytoplasmic domain. The ECD contains an N-terminal globular domain, a cysteine-rich domain, and two fibronectin type III domains (5). The cytoplasmic domain contains a juxtamembrane motif with two tyrosine residues, which are the major autophosphorylation sites, a kinase domain, and a sterile alpha motif (SAM) (5). The ECD of human EphA2 shares 90-94% aa sequence identity with mouse, bovine, and canine EphA2, and approximately 45% aa sequence identity with human EphA1, 3, 4, 5, 7, and 8. EphA2 becomes autophosphorylated following ligand binding (6, 7) and then interacts with SH2 domain-containing PI3-kinase to activate MAPK pathways (8, 9). Reverse signaling is also propagated through the Ephrin ligand. Transcription of EphA2 is dependent on the expression of E-Cadherin (10), and can be induced by p53 family transcription factors (11). EphA2 is upregulated in breast, prostate, and colon cancer vascular endothelium. Its ligand, EphrinA1, is expressed by the local tumor cells (12, 13). In some cases, EphA2 and EphrinA1 are expressed on the same blood vessels (14). EphA2 signaling cooperates with VEGF receptor signaling in promoting endothelial cell migration (13). The gene encoding human EphA2 maps to a region on chromosome 1 which is frequently deleted in neuroectodermal tumors (15).

  • References:
    1. Poliakov, A. et al. (2004) Dev. Cell 7:465.
    2. Surawska, H. et al. (2004) Cytokine Growth Factor Rev. 15:419. 
    3. Pasquale, E.B. (2005) Nat. Rev. Mol. Cell Biol. 6:462.
    4. Davy, A. and P. Soriano (2005) Dev. Dyn. 232:1.
    5. Bohme, B et al. (1993) Oncogene 8:2857.
    6. Pandey, A. et al. (1995) Science 268:567.
    7. Bartley, T.D. et al. (1994) Nature 368:558.
    8. Pandey, A. et al. (1994) J. Biol. Chem. 269:30154.
    9. Miao, H. et al. (2001) Nat. Cell Biol. 3:527.
    10. Orsulic, S. and R. Kemler (2000) J. Cell Sci. 113:1793.
    11. Dohn, M. et al. (2001) Oncogene 20:6503.
    12. Zelinski, D.P. et al. (2001) Cancer Res. 61:2301.
    13. Brantley, D.M. et al. (2002) Oncogene 21:7011.
    14. Ogawa, K. et al. (2000) Oncogene 19:6043.
    15. Sulman, E.P. et al. (1997) Genomics 40:371.
  • Entrez Gene IDs:
    1969 (Human); 13836 (Mouse)
  • Alternate Names:
    ARCC2; EC 2.7.10; EC 2.7.10.1; Eck; ECKepithelial cell receptor protein tyrosine kinase; EPH receptor A2; EphA2; ephrin type-A receptor 2; Epithelial cell kinase; Myk2; Sek2; soluble EPHA2 variant 1; Tyrosine-protein kinase receptor ECK
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Alteration of the EphA2/Ephrin-A signaling axis in psoriatic epidermis.
    Authors: Gordon K, Kochkodan J, Blatt H, Lin S, Kaplan N, Johnston A, Swindell W, Hoover P, Schlosser B, Elder J, Gudjonsson J, Getsios S
    J Invest Dermatol, 2013;133(3):712-22.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  2. EphA2 cleavage by MT1-MMP triggers single cancer cell invasion via homotypic cell repulsion.
    Authors: Sugiyama N, Gucciardo E, Tatti O, Varjosalo M, Hyytiainen M, Gstaiger M, Lehti K
    J Cell Biol, 2013;201(3):467-84.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. A novel extracellular Hsp90 mediated co-receptor function for LRP1 regulates EphA2 dependent glioblastoma cell invasion.
    Authors: Gopal U, Bohonowych JE, Lema-Tome C
    PLoS ONE, 2011;6(3):e17649.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  4. Protein kinase A can block EphA2 receptor-mediated cell repulsion by increasing EphA2 S897 phosphorylation.
    Authors: Barquilla A, Lamberto I, Heynen-Genel S, Brill L, Pasquale E
    Mol Biol Cell, 0;27(17):2757-70.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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