Fig. 3
Retrieval of recombinant α-globin fusion protein variants purified by affinity Ni2+-NTA chromatography under denaturing conditions and analyzed by 15% SDS − PAGE and Coomassie Blue staining: A SDS-PAGE analysis of the purification of 10xHis-XaSITE-α-globin-HA protein (~ 18.7 kDa). Lane 1: Crude extract of induced bacterial cells transformed by pET-16b-α globin-HA; Lane 2: Flow-through fraction of the Ni2+-NTA affinity column; Lane 3: Wash Buffer extract with 10 mM imidazole; Lane 4: Elution with 20 mM imidazole; Lane 5: Elution with 40 mM imidazole; Lane 6: protein molecular mass marker; Lane 7: Elution with 100 mM imidazole; Lane 8: Elution with 200 mM imidazole; Lane 9: Elution with 400 mM imidazole. B SDS-PAGE analysis of the purification of 10xHis-XaSITE-TAT-α-globin-HA protein (~ 20.2 kDa). Lane 1: Crude extract of induced bacterial cells transformed by pET-16b-TAT-α globin-HA; Lane 2: Flow-through fraction of the Ni2+-NTA affinity column; Lane 3: Wash Buffer extract with 10 mM imidazole; Lane 4: Elution with 20 mM imidazole; Lane 5: Elution with 40 mM imidazole; Lane 6: Elution with 100 mM imidazole; Lane 7: Elution with 200 mM imidazole; Lane 8: Elution with 400 mM imidazole; Lane 9: protein molecular mass marker