Figure 2

Coomassie blue staining showing the purification process of (His)6-p53 fusion protein using nickel chelated affinity chromatography. 15 μl of each fraction were resolved through 10% SDS-PAGE, followed by Coomassie blue staining. The molecular weight of the purified protein was ~53 kDa. M, molecular weight marker; lane 1, bacterial cell lysate containing (His)6-p53 fusion protein; lane 2, flow-through fraction of the nickel chelated affinity chromatography; lane 3, washed fraction with binding buffer, lane 4–7, washed fractions with washing buffer containing increasing concentration of imidazole (20, 60, 40 and 80 mM, respectively), lane 8, the purified (His)6-p53 fusion protein obtained after eluting with 1 M imidazole