Human/Rat EphA5 Antibody

Catalog #: MAB541
3 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB541
MAB541-SP

Detection of EphA5 in U‑118‑MG Human Cell Line by Flow Cytometry.

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All EphA5 Antibodies

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Citations (3)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Rat EphA5 Antibody Summary

Species Reactivity

Human, Rat

Specificity

Detects rat EphA5 in direct ELISAs and Western blots. In direct ELISAs, 5‑15% cross reactivity with recombinant mouse (rm) EphA4 and rmEpha8 is observed and no cross-reactivity with recombinant human (rh) EphA1, rmEphA2, -A3, -A6, -A7, recombinant rat EphB1, -B2, or rmEphB3 is observed.

Source

Monoclonal Mouse IgG₁ Clone # 86731

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat EphA5
Ser58-Gln573 (Asp170Glu)
Accession # P54757

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

Recombinant Rat EphA5 Fc Chimera (Catalog # 541-A5)

Flow Cytometry

0.25 µg/10⁶ cells

See below

CyTOF-ready

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

Immunocytochemistry

8-25 µg/mL

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.

Scientific Data

Flow Cytometry

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Detection of EphA5 in U‑118‑MG Human Cell Line by Flow Cytometry. U-118-MG human glioblastoma/astrocytoma cell line was stained with Mouse Anti-Human/Rat EphA5 Monoclonal Antibody (Catalog # MAB541, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry

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EphA5 in U-118-MG Human Cell Line. EphA5 was detected in immersion fixed U-118-MG human glioblastoma/astrocytoma cell line using 10 µg/mL Mouse Anti-Human/Rat EphA5 Monoclonal Antibody (Catalog # MAB541) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

Background: EphA5

EphA5, also known as Ehk1, Bsk, Cek7, Hek7, and Rek7 (1), is a member of the Eph receptor family which binds members of the ephrin ligand family. There are two classes of receptors, designated A and B. Both the A and B class receptors have an extracellular region consisting of a globular domain, a cysteine-rich domain, and two fibronectin type III domains. This is followed by the transmembrane region and cytoplasmic region. The cytoplasmic region contains a juxtamembrane motif with two tyrosine residues, which are the major autophosphorylation sites, a kinase domain, and a conserved sterile alpha motif (SAM) in the carboxy tail which contains one conserved tyrosine residue. Activation of kinase activity occurs after ligand recognition and binding. EphA5 has been shown to bind ephrin-A5, ephrin-A1,
ephrin‑A2, ephrin-A3, and ephrin-A4 (2, 3). The extracellular domains of rat EphA5 share 98.5% amino acid identity with the mouse homolog and 96.5% identity with the human homolog. Only membrane-bound or Fc-clustered ligands are capable of activating the receptor in vitro. While soluble monomeric ligands bind the receptor, they do not induce receptor autophosphorylation and activation (2). In vivo, the ligands and receptors display reciprocal expression (3). It has been found that nearly all receptors and ligands are expressed in developing and adult neural tissue (3). The Eph/ephrin families also appear to play a role in angiogenesis (3).

References

Eph Nomenclature Committee [letter] (1997) Cell 90:403.
Flanagan, J.G. and P. Vanderhaeghen (1998) Annu. Rev. Neurosci. 21:309.
Pasquale, E.B. (1997) Curr. Opin. Cell Biol. 9:608.

Entrez Gene IDs

2044 (Human); 13839 (Mouse); 79208 (Rat)

Alternate Names

Bsk; Cek7; EC 2.7.10; EC 2.7.10.1; Ehk1; EHK-1; EHK1Hek7; EK7; EPH homology kinase 1; Eph homology kinase-1; EPH receptor A5; EphA5; EPH-like kinase 7; ephrin type-A receptor 5; Hek7; receptor protein-tyrosine kinase HEK7; Rek7; TYRO4HEK7CEK7; tyrosine-protein kinase receptor EHK-1

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Citations for Human/Rat EphA5 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.

3 Citations: Showing 1 - 3
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