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Mouse EphA1 Antibody

Catalog #: AF3034 Datasheet
Catalog # Availability Size / Price Qty
AF3034
AF3034-SP
Detection of Mouse EphA1 by Western Blot.
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Mouse EphA1 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse EphA1 in direct ELISAs and Western blots. In direct ELISAs, approximately 35% cross-reactivity with recombinant human EphA1 and less than 5% cross-reactivity with recombinant mouse (rm) EphA4, rmEphA6, and rmEphA7 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse EphA1
Glu27-Glu548
Accession # AAH71215
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.

Applications

Recommended Concentration
Sample
Western Blot
0.25 µg/mL
See below
Flow Cytometry
2.5 µg/106 cells
D3 mouse embryonic stem cell line
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.
 

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.

Scientific Data

Western Blot Detection of Mouse EphA1 antibody by Western Blot. View Larger

Detection of Mouse EphA1 by Western Blot. Western blot shows lysates of mouse lung tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse EphA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3034) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for EphA1 at approximately 113 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry EphA1 antibody in Mouse Thymus by Immunohistochemistry (IHC-Fr). View Larger

EphA1 in Mouse Thymus. EphA1 was detected in perfusion fixed frozen sections of mouse thymus using Goat Anti-Mouse EphA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3034) at 1.7 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 thymic stromal cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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.

Background: EphA1

EphA1, also known as Eph and Esk, is a member of the Eph receptor tyrosine kinase family and binds several Ephrin-A ligands. The A and B class Eph proteins share a common structural organization (1-4). The mouse EphA1 cDNA encodes a 977 amino acid (aa) precursor that includes a 24 aa signal sequence, a 524 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 408 aa cytoplasmic domain. The ECD contains an N-terminal globular domain, a cysteine-rich domain, and two fibronectin type III domains. The cytoplasmic domain contains a juxtamembrane motif with two tyrosine residues which are the major autophosphorylation sites, a kinase domain, and a conserved sterile alpha motif (SAM) (5, 6). Within the ECD, mouse EphA1 shares 84% aa sequence identity with human EphA1, approximately 40% aa sequence identity with mouse EphA2, 3, 4, 6, 7, and 8, and 27% aa sequence identity with mouse EphA5. A splice variant of mouse EphA1 lacks the transmembrane segment and is predicted to exist as a soluble molecule (7). Membrane bound or clustered Ephrin ligands interact with EphA1 and activate its kinase domain which is capable of Ser, Thr, and Tyr phosphorylation (7). Reverse signaling is propagated through the Ephrin ligand. EphA1 is widely expressed in differentiated epithelial cells, particularly in bone marrow, spleen, thymus, and testes (5, 7). It is expressed on CD4+ T cells but not on CD19+ B cells (8). On T cells, EphA1 induces Tyr phosphorylation of PYK2 and promotes chemokine-induced chemotaxis (8). EphA1 is upregulated or downregulated in a variety of human carcinomas and is implicated in tumor invasiveness (2, 9, 10).

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. Coulthard, M.G. et al. (2001) Growth Factors 18:303.
  6. Lickliter, J.D. et al. (1996) Proc. Natl. Acad. Sci. USA 93:145.
  7. Douville, E.M.J. et al. (1992) Mol. Cell. Biol. 12:2681.
  8. Aasheim, H-C. et al. (2005) Blood 105:2869.
  9. Iwase, T. et al. (1993) Biochim. Biophys. Res. Commun. 194:698.
  10. Hirai, H. et al. (1987) Science 238:1717.
Entrez Gene IDs
2041 (Human); 13835 (Mouse)
Alternate Names
EC 2.7.10; EC 2.7.10.1; EPH receptor A1; eph tyrosine kinase 1; EphA1; EPHephrin type-A receptor 1; EPHT; EPHT1erythropoietin-producing hepatoma amplified sequence; Esk; MGC163163; oncogene EPH; soluble EPHA1 variant 1; soluble EPHA1 variant 2; Tyrosine-protein kinase receptor EPH

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