FIGURE 1.

MT digestion. Panel A shows an SDS-PAGE analysis of native MTs and subtilisin dMTs. Undigested α- and β-subunits of tubulin do not separate under standard conditions. Digestion, however, leads to clear separation of the subunits. We confirmed complete digestion of β-tubulin using a Western blot with monoclonal anti-β-tubulin antibodies. Panel B shows the Ponceau S-stained blot, and panel C the Western blot after detection of the primary antibody with a secondary, alkaline phosphatase-conjugated antibody and incubation with NBT/BCIP as substrate.