Mouse Anti-EREG Recombinant Antibody (clone 9E5) (CAT#: PABL-084)

Figure 1 Decreased vinculin and paxillin accumulation in focal adhesion by anti-EREG antibody (9E5) in DLD1 cells.

DLD-1 (a, b, e, f, g, h) or AGS (c, d) cells were incubated with control IgG (a, c, e, g) or 9E5 IgG (b, d, f, h) at 37 °C for 30 min. Cells were fixed and labeled with vinculin (a–d), paxillin (e, f), or integrin β5 (g, h). Arrows shows focal adhesion; bars, 5 μm. B. The effects of anti-EREG antibody (9E5) on cell spreading. For cell spreading assay, cells were treated with control IgG (a, c, e) or 9E5 IgG (b, d, f) in suspension, plated on vitronectin-coated coverslips, and incubated at 37 °C for 15 min (a, b) or 90 min (c, d, e, f). Cells were washed, fixed, and visualized by staining with Alexa 594-conjugated phalloidin (a, b, c, d: red) or integrin β5 (e, f: green); bars, 10 μm.

Iijima, M., Anai, M., Kodama, T., & Shibasaki, Y. (2017). Epiregulin-blocking antibody inhibits epiregulin-dependent EGFR signaling. Biochemical and biophysical research communications, 489(1), 83-88.

Figure 2 Effect of anti-EREG antibody on EGFR signaling.

DLD1 cells were incubated with 9E5 IgG for 30 min and treated with EGF or EREG for 5 min. Cells were immunoblotted with anti-p-EGFR, p-Erk, or p-Akt antibodies.

Iijima, M., Anai, M., Kodama, T., & Shibasaki, Y. (2017). Epiregulin-blocking antibody inhibits epiregulin-dependent EGFR signaling. Biochemical and biophysical research communications, 489(1), 83-88.

Figure 3 Immunofluorescent staining of antibodies in human colorectal carcinoma cells.

DLD-1 and HCT116 cells, representing two human colorectal carcinoma cell lines, were fixed with paraformaldehyde and treated with or without 0.2% Triton X-100 solution (localization to endomembranes was observed; white arrow).

Lee, Y. H., Iijima, M., Kado, Y., Mizohata, E., Inoue, T., Sugiyama, A.,... & Kodama, T. (2013). Construction and characterization of functional anti-epiregulin humanized monoclonal antibodies. Biochemical and biophysical research communications, 441(4), 1011-1017.

Figure 4 Flow cytometry analysis.

Antibodies were tested for their ability bind to the surface of each cell line (DLD-1, HCT116, and AGS). Green line: 0.1 lg/mL, blue line: 0.5 lg/mL, purple line: 1 lg/mL, red line: negative control (0 lg/mL).

Lee, Y. H., Iijima, M., Kado, Y., Mizohata, E., Inoue, T., Sugiyama, A.,... & Kodama, T. (2013). Construction and characterization of functional anti-epiregulin humanized monoclonal antibodies. Biochemical and biophysical research communications, 441(4), 1011-1017.

Figure 5 Single-cycle kinetic analysis of 9E5, HM0, and HM1 antibody binding to an amine-coupled CM5 chip on Biacore.

hEpiregulin-Fc was injected at concentrations of 1.5 lM, 300, 60, 12, and 2.4 nM, respectively, and analyzed in duplicate binding cycles over three differing capacity reaction surfaces.

Lee, Y. H., Iijima, M., Kado, Y., Mizohata, E., Inoue, T., Sugiyama, A.,... & Kodama, T. (2013). Construction and characterization of functional anti-epiregulin humanized monoclonal antibodies. Biochemical and biophysical research communications, 441(4), 1011-1017.

Figure 1 Mouse Anti-EREG Recombinant Antibody (PABL-084) in SDS-PAGE

SDS-PAGE analysis of PABL-084 in non-reduced (Lane 1, 2 μg) and reduced (Lane 2, 2 μg) conditions.

Figure 2 Mouse Anti-EREG Recombinant Antibody (PABL-084) in HPLC

The purity of PABL-084 was greater than 98% as determined by SEC-HPLC.