Human EphA2 Antibody

Catalog # Availability Size / Price Qty
AF3035
AF3035-SP
Detection  of Human EphA2 by Western Blot.
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Product Details
Citations (6)
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Human EphA2 Antibody Summary

Species Reactivity
Human
Specificity
Detects human EphA2 in direct ELISAs and Western blots. In direct ELISAs, less than 45% cross-reactivity with recombinant mouse 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

Western Blot Detection  of Human EphA2 by Western Blot. View Larger

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.

Flow Cytometry Detection  of EphA2 in A431 Human Cell Line by Flow Cytometry. View Larger

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. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

Citations for Human EphA2 Antibody

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.

6 Citations: Showing 1 - 6
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  1. Brucine Suppresses Vasculogenic Mimicry in Human Triple-Negative Breast Cancer Cell Line MDA-MB-231
    Authors: MR Xu, PF Wei, MZ Suo, Y Hu, W Ding, L Su, YD Zhu, WJ Song, GH Tang, M Zhang, P Li
    Biomed Res Int, 2019;2019(0):6543230.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. Engineered nanointerfaces for microfluidic isolation and molecular profiling of tumor-specific extracellular vesicles
    Authors: E Reátegui, KE van der Vo, CP Lai, M Zeinali, NA Atai, B Aldikacti, FP Floyd, A H Khankhel, V Thapar, FH Hochberg, LV Sequist, BV Nahed, B S Carter, M Toner, L Balaj, D T Ting, XO Breakefiel, SL Stott
    Nat Commun, 2018;9(1):175.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  3. 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 Types: Whole Tissue
    Applications: IHC-Fr
  4. 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 Types: Cell Lysates
    Applications: Western Blot
  5. 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 Types: Whole Cells
    Applications: Flow Cytometry
  6. 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 Types: Cell Lysates
    Applications: Western Blot

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