Figure 3. Analysis of the purity of BoNT/A HCN, Hcc and LC-HN domains by SDS-PAGE.
Coomassie-stained SDS-PAGE gels were run to characterize the purity of domain fractions. Molecular weight markers (MW) are in the left lane of each gel. Lanes 1–3 HCC (Lane 1, HCC after refolding from denatured inclusion bodies; Lane 2, excluded elution shoulder fractions showing prominent “ladder” containing improperly oxidized HCC species identified by RP-HPLC (see Figure 5); Lane 3, HCC main peak). Lanes 4–6 HCN (Lane 4, clarified HCN lysate; Lane 5, partially purified HCN ion exchange pool; Lane 6, Ni-NTA IMAC pool). Lanes 7–8 LC-HN (Lane 7, partially purified ion exchange pool; Lane 8, purified LC-HN pool).