Human Ephrin-B3 Antibody

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.

Detection of Ephrin-B3 by Western Blot

EphrinB3 binds and activates EphA4, EphB1 and EphB6 receptors in SC. a, c, d Orthogonal views of SC activation with clustered EphrinB3 shows that EphrinB3 binds EphB6 (a), EphA4 (c) and EphB1 (d) receptors on SC. b Western blot confirming the presence of these receptors in SC. e, f Bound EphrinB3 activation of EphA4 and EphB receptors viewed by immunodetection of phosphorylated forms. g, h Western blot illustrating that SC incubation with myelin increased the phosphorylation of EphA4 (g) and EphB1 + B2 (h) which was not the case when myelin was previously blocked by anti-EphrinB3 (O.D. p-EphB1 + B2/GAPDH: one-way ANOVA p = 0.003, F(2,15) = 8.57, followed by a Tukey’s multiple comparison test) and O.D. of p-EphA4/GAPDH: one-way ANOVA p = 0.0036, F(2,15) = 8.44, followed by a Tukey’s multiple comparison test). Data are expressed as ratio of the optical density (O.D.) of the bands (mean values ± SD) from three independent experiments, control (n = 6), myelin (n = 6), myelin + anti-EphrinB3 (n = 6). Scale bar 20 µm Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31011859), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Ephrin-B3 by Western Blot

EphrinB3 improves SC adhesion and spreading on FN-coated surfaces. a Quantification of the number of adhered and polarized SC seeded on surfaces coated with EphrinB3 with or without FN. Data are expressed as ratio respect to Fc-coated surfaces. EphrinB3 (10 µg/mL), EphrinB3 (10 µg/mL) + FN (2 µg/cm2), n = 7 per group. b1 Diagram of the adhesion assay on FN. Coverslips divided by silicon strips were coated with Fc + FN (control) or EphrinB3 + FN in each half. After strip removal, cells pre-incubated with IgG or anti-Integrin beta 1 were seeded homogenously on the coverslip. b2 Blocking Integrin beta 1 decreased SC adhesion and polarization on EphrinB3 + FN compared with Fc + FN. Arrows show examples of “round cells”, scale bar 200 µm. c Quantification of SC adhesion and polarization on EphrinB3 when SC are pre-incubated with IgG or anti-integrin beta 1, n = 15–17 per group. d–e Western blot shows an increased expression of Integrin beta 1 when SC are activated by myelin extracts. One-way ANOVA p = 0.011, F(2,12) = 6.62, followed by Tukey’s multiple comparisons test p < 0.05, n = 5 per group. Data are expressed as mean values ± SD. In a, c*/** are used for comparison of a group with its hypothetical mean of 1 by one-sample two-tailed t-test, and #/## for comparison between two different groups by two-tailed Mann–Whitney test.*/#p < 0.05; **/##p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31011859), licensed under a CC-BY license. Not internally tested by R&D Systems.