FIGURE 4.

IMAC purification of the Cysta-tag–α₁AT fusion. Cysta-tag and α₁AT separated by the factor X_a-cleavable sequence were transiently expressed in N. benthamiana leaves. (A) α₁AT recovery (line) and purity (columns) rates during purification. Calculations were made based on ELISA assays for α₁AT and leaf total soluble protein determinations. Data are mean values of three independent purification rounds ±SD. (B) Coomassie blue-stained polyacrylamide gels following SDS-PAGE showing key protein fractions during the purification process. Identity of the Cysta-tag–α₁AT fusion was confirmed by immunodetections with anti-α₁AT and anti-SlCYS8 primary antibodies (C). Arrowheads point to the Cysta-tag–α₁AT fusion, closed circles to free Cysta-tag and asterisk (^∗) to free α₁AT.