Detection of Mouse and Rat Ephrin‑B3 by Western Blot.
Western blot shows lysates of mouse brain tissue and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Ephrin‑B3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF395) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Ephrin‑B3 at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
Ephrin-B3, also known as NLERK-2, Elk-L3, EFL-6, ELF-3 and LERK-8 (1), is a member of the Ephrin ligand family which binds members of the Eph receptor family. All ligands share a conserved extracellular sequence, which most likely corresponds to the receptor binding domain. This conserved sequence consists of approximately 125 amino acids (aa) and includes four invariant cysteines. The B-class ligands are transmembrane proteins which can be tyrosine phosphorylated upon receptor ligation. The cytoplasmic domains are approximately 80 aa long and are highly conserved, especially the last 33 aa. Several signaling molecules have been shown to interact with the cytoplasmic region, although specific signaling roles have yet to be elucidated. Ephrin-B3 has been shown to bind EphA4, EphB1, EphB2, and EphB3 (2, 3). The extracellular domains of murine and human Ephrin-B3 share 98% aa identity. 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 the receptors and ligands are expressed in developing and adult neural tissue (3). The Ephrin/Eph families also appear to play a role in angiogenesis (3).
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