Anti-Human EPHB4 Recombinant Antibody (MOR03640) (CAT#: TAB-001WM)

Figure 1 The specificity of Fabs for EphB4 and cross-reactivity to both human and murine EphB4 in ELISA.

Anti-EphB4 Fab fragments (purified, MH-tagged) were added (100 uL/well, 100 g/mL final concentration in chemiblock) to respective wells and incubated for 1 h at RT.Then 100 ul mEphrinB2-Fc fusion protein either spiked with the respective anti-EphB4 Fab or with PBS was added (25 ng/ml mEphrinB2+/-100 ug/ml Fab diluted in chemiblock)and incubated for max. 20 min at RT. The amount of bound EphrinB2 was detected using biotinylated goat anti-EphrinB2 antibody (0.05 g/ml final concentration diluted in chemiblock, 1 h at RT), AP-conjugated streptavidin and AttoPhos substrate.

Figure 2 The FACS analysis of MCF-7 breast cancer cell line with anti-EphB4 Fabs.

FACS analysis was used to confirm that the MCF-7 breast cancer cell line is highly positive for EphB4 expres sion. EphB4 Fabs were incubated at a concentration of 10 ug/mL with MCF-7 cells at 4°C. for 1 hour. Afterwashing with PBS, the cells were incubated with FITC-labelled sec ondary antibody. After a final PBS wash, the cell-associated fluorescence was measured using a Becton Dickinson Calibur Fluorescence activated cell sorter.