Characterization of Human Colorectal Cancer MDR1/P-gp Fab Antibody
Figure 5
Expression, purification, and verification of the P-gp21. (a) SDS-PAGE image of the P-gp21 expression induced by IPTG: before induction (lane 1), induced for 1 hour (lane 2), induced for 2 hours (lane 3), induced for 3 hour (lane 4), and induced for 4 hours (lane 5); (b) purification of the P-gp21 protein by HiTrap HP, before passing Ni affinity column (lane 1), sample after passing Ni affinity column (lane 2), the washout of the Ni affinity column with 50 mM imidazole (lane 3), the washout of the Ni affinity column with 150 mM imidazole (lane 4), and the washout of the Ni affinity column with 300 mM imidazole (lane 5); (c) P-gp21 protein verified by Western blot: the recombinant with P-gp21 before induction (lane 1), purified P-gp21 protein (lane 2), antigen (P-gp21 protein), primary antibody (mouse anti-His antibody), and secondary antibody (HRP-labeled goat anti-mouse IgG antibody).
