Recombinant Human EFNB2 protein(Met1-Ala229), His-tagged

• Cat.No.: EFNB2-3191H
• Product Overview: Recombinant Human EFNB2 (NP_004084.1) (Met 1-Ala 229) was expressed in HEK293, fused with a C-terminal polyhistidine tag.

Specification

• Species: Human
• Source: HEK293
• Tag: His
• Protein Length: 1-229 a.a.
• Form: Lyophilized from sterile PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Bio-activity: Measured by its binding ability in a functional ELISA. Immobilized human EFNB2 at 2 μg/ml (100 μl/well) can bind human EphB4 with a linear range of 1.56-12.5 ng/ml.
• Molecular Mass: The recombinant human EFNB2 consists of 213 amino acids after removal of the signal peptide and predicts a molecular mass of 23.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh EFNB2 is approximately 35-40 kDa due to glycosylation.
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method
• Purity: > 97 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: EFNB2 ephrin-B2 [ Homo sapiens ]
• Official Symbol: EFNB2
• Synonyms: EFNB2; ephrin-B2; EPLG5; eph related receptor tyrosine kinase ligand 5; HTK ligand; Htk L; HTKL; LERK5; ligand of eph related kinase 5; MGC126226; MGC126227; MGC126228; LERK-5; ligand of eph-related kinase 5; eph-related receptor tyrosine kinase ligand 5; Htk-L
• Gene ID: 1948
• mRNA Refseq: NM_004093
• Protein Refseq: NP_004084
• MIM: 600527
• UniProt ID: P52799

Case Study

Case 1: Zhang Q, et al. Arch Biochem Biophys. 2024

EFNB2 downregulation exacerbates intervertebral disc degeneration by promoting NP cell apoptosis via Bax/Bcl-2 imbalance and caspase-3 activation. Restoring EFNB2 activates PI3K/AKT survival signaling and inhibits ERK phosphorylation, mitigating IL-1β-induced disc degeneration, positioning it as a therapeutic target for spinal health and disc degeneration therapies.

Fig1. EFNB2 protein expression were examined in collected NP tissues using Immunoblotting.

Fig2. NP cells were transduced with EFNB2 OE, treated or non-treated with IL-1β, and examined for the protein levels of p-ERK1/2 and ERK1/2.

Case 2: Wang W, et al. Stem Cell Res Ther. 2020

EphrinB2 overexpression enhances DPSCs’ osteogenic differentiation via EphB4 reverse signaling, boosting alveolar bone regeneration in scaffold-based therapies. In vitro and canine models confirm increased trabecular bone volume, positioning ephrinB2-engineered DPSCs as a breakthrough for bone defect repair and dental regenerative medicine.

Fig1. Expression of p-ephrinB2, ephrinB2, p-EphB4, and EphB4 in hDPSCs during osteogenic differentiation.

Fig2. Western blot analysis confirmed that ephrinB2 reverse signaling was activated by 1 μg/ml EphB4-Fc.

Application

1. Therapeutic Potential of Recombinant EFNB2 Protein in Bone Regeneration and Spinal Repair Recombinant ephrin-B2 (EFNB2) protein, a key regulator of EphB4-mediated reverse signaling, offers transformative potential in regenerative medicine. By enhancing osteogenic differentiation of dental pulp stem cells (DPSCs) and promoting bone matrix deposition, EFNB2 accelerates alveolar bone defect repair and mitigates intervertebral disc degeneration (IDD). Its dual role in activating PI3K/AKT survival pathways and suppressing ERK-driven apoptosis positions it as a therapeutic cornerstone for bone loss and spinal disorders. 2. Mechanistic Insights and Preclinical Validation In vitro studies demonstrate recombinant EFNB2 activates EphB4 reverse signaling, upregulating osteogenic markers (e.g., Runx2, OCN) and stabilizing β-catenin to drive DPSC mineralization. Preclinical models, including canine alveolar defects and rodent IDD systems, confirm EFNB2-scaffold composites (e.g., PuraMatrix) enhance trabecular bone volume (BV/TV) and reduce NP cell apoptosis. Dose-dependent EFNB2 delivery restores disc hydration and ECM synthesis, counteracting IL-1β-induced degeneration via PI3K/AKT/GSK-3β pathway modulation. 3. Challenges and Innovations in Clinical Translation Current limitations include short protein half-life and off-target Eph receptor activation. Advances in sustained-release hydrogels, CRISPR-engineered DPSCs, or fusion proteins (EFNB2-Fc) improve localized delivery and signaling specificity. Combining EFNB2 with BMP-2 or anti-inflammatory agents may synergize bone formation and reduce fibrosis. Ongoing research explores its utility in osteoporosis and craniofacial reconstruction, solidifying EFNB2 as a multi-target therapy for musculoskeletal and dental regeneration.

Fig1. Molecular compartmentalization of ephrinB2 and EphB4. (Andras Piffko, 2022)