Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Recombinant Protein, Yeast

Applications

Validated:

Knockout Validated, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, ELISA Capture

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 371805
View all EphA2 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human EphA2
Gln25-Asn534
Accession # P29317

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human EphA2 Antibody

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

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

A431 human epithelial carcinoma cell line was stained with Mouse Anti-Human EphA2 Monoclonal Antibody (Catalog # MAB3035, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.
EphA2 antibody in A431 Human Cell Line by Immunocytochemistry (ICC).

EphA2 in A431 Human Cell Line.

EphA2 was detected in immersion fixed A431 human epithelial carcinoma cell line using Mouse Anti-Human EphA2 Monoclonal Antibody (Catalog # MAB3035) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI(blue). Specific staining was localized to the cell surface. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
EphA2 Antibody Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

EphA2 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

EphA2 was detected in immersion fixed A431 human epithelial carcinoma cell line but is not detected in EphA2 knockout (KO) A431 Human Cell Line cell line using Mouse Anti-Human EphA2 Monoclonal Antibody (Catalog # MAB3035) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; Catalog # NL009) and counterstained with DAPI (blue). Specific staining was localized to cell membranes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
EphA2 in PC-3 Human Prostate Cancer Cell Line.

EphA2 in PC-3 Human Prostate Cancer Cell Line.

Fixed PC-3 cell stained with MAB3035 and detected with a secondary anti-mouse AlexaFluor488-conjugated antibody. Left: wildtype PC-3 cells (EphA2 expressing); right: PC-3 cells EphA2 Knock-out. Image from a verified customer review.
Detection of EphA2 by Flow Cytometry

Detection of EphA2 by Flow Cytometry

Identification of non-competing anti-EphA2 mAbs.(A) Histograms for EphA2-positive cell lines HEC-1-A and PC-3 first stained with unlabeled EphA2 mAb 1C1 or 3035, an IgG control, or buffer only, followed by staining with either labeled 1C1 (1C1-A488) or 3035 (3035-A488). A488 fluorescence is detected by conducting flow cytometry on 10,000 live, single cells. (B) Percent staining by 1C1-A488 or 3035-A488 relative to the HEC-1-A or PC-3 samples first incubated with buffer only. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25894652), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human EphA2 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: A431 human epithelial carcinoma cell line

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line

Knockout Validated

EphA2 was detected in immersion fixed A431 human cell line but is not detected in EphA2 knockout (KO) A431 human cell line.

Reviewed Applications

Read 4 reviews rated 4.5 using MAB3035 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

Alternate Names

Eck, Myk2, Sek2

Entrez Gene IDs

1969 (Human); 13836 (Mouse)

Gene Symbol

EPHA2

UniProt

P29317

Additional EphA2 Products

Product Documents for Human EphA2 Antibody

Certificate of Analysis

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Product Specific Notices for Human EphA2 Antibody

For research use only

Citations for Human EphA2 Antibody

Customer Reviews for Human EphA2 Antibody (4)

4.5 out of 5
4 Customer Ratings
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Immunocytochemistry
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: PC-3 human prostate cancer cell line
    Species: Human
    Verified Customer | Posted 08/19/2025
    Fixed PC-3 cell stained with MAB3035 and detected with a secondary anti-mouse AlexaFluor488-conjugated antibody. Left: wildtype PC-3 cells (EphA2 expressing); right: PC-3 cells EphA2 Knock-out.
    Cells were fixed with 4% PFA for 20 minutes at room temperature and then permeabilised with Triton X-100 0.2%. The cells were stained with MAB3035 diluted 1:200 in PBS (containing 3% BSA) for 1 hour at room temperature. The antibody was then detected using a fluorescent anti-mouse secondary antibody.
    Human EphA2 Antibody MAB3035
  • Human EphA2 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 07/05/2022
    Human EphA2 Antibody MAB3035
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 21245385
    Species: Human
    Verified Customer | Posted 02/19/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 24036371
    Species: Other
    Verified Customer | Posted 02/19/2015

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