Human EphA2 Alexa Fluor® 647-conjugated Antibody

Catalog # Availability Size / Price Qty
FAB3035R-100UG
Detection of EphA2 in A431 Human Cell Line by Flow Cytometry.
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Human EphA2 Alexa Fluor® 647-conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects human EphA2 in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant mouse EphA4, A5, A6, A7, A8, or recombinant rat EphB1 is observed.
Source
Monoclonal Mouse IgG2A Clone # 371805
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human EphA2
Gln25-Asn534
Accession # P29317
Formulation
Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 647

Applications

Recommended Concentration
Sample
Flow Cytometry
0.25-1 µg/106 cells
See below

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 Example

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 Mouse Anti-Human EphA2 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # FAB3035R, filled histogram) or isotype control antibody (Catalog # IC003R, open histogram). View our protocol for Staining Membrane-associated Proteins.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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, 2, 3, 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

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