Human Anti-EPHA4 Recombinant Antibody (clone m119) (CAT#: HPAB-0006-DJB)

Figure 1 Binding of IgG1s E1 (m101), E2, E5 (m105), and E19 (m119) to mouse and human EphA4 measured by ELISA.

Serially diluted IgG1s were added to EphA4-coated wells. HRP-conjugated anti-FLAG Ab was used for detection.

Figure 2 Binding of IgG1s m101, m105, and m119 to mouse and human EphA4 measured by ELISA.

Serially diluted IgG1s were added to EphA4-coated wells. HRP-conjugated anti-Fab Ab was used for detection.

Figure 3 IgG1s m101, m105, and m119 recognize human EphA4 ligand binding domain (LBD).

The EphA4 LBD recombinant proteins were purified and subjected to western blot analysis using the indicated IgG1s.

Figure 4 IgG1s m101, m105, and m119 exhibit strong binding affinity towards the EphA4 LBD.

Isothermal titration calorimetry (ITC) experiments were carried out by incubating 20 MM Eph LBD protein in the cell with 1 MM antibodies. (A) Human IgG showed no binding; (B) m101 showed binding affinity with Kd=˜83.0 nM; (C) m105 showed strong binding affinity with Kd=˜8.6 nM; (D) m119 showed binding affinity with Kd=˜16.8 nM.

Figure 5 IgG1s m101, m105, and m119 block the ephrin-A1/EphA4 interaction.

The plot indicates that the 3 IgG1s were able to block the interaction between mouse EphA4 extracellular domain with mouse ephrin-A1, at a nano molar range (IC50: m101=0.71 nM; m105=0.35 nM; and m119=1.81 nM).

Figure 6 IgG1 m119 induces the tyrosine phosphorylation of EphA4 in cultured cortical neurons.

DIV 7 cortical neurons were treated with 2 mg/ml of m101 (101), m105 (105), m119 (119), and ephrin-A1 (A1) as positive control. The cells were harvested and subjected to immunoprecipitation and western blot analysis using the indicated antibodies.