The insert was cloned into the SpyTag/SpyCatcher cassette using the method previously described10 to make pET28a SpyTag-PhyC- SpyCatcher (GenBank accession number KU608330) and pET28a SpyTag DA-PhyC-SpyCatcher. pET28a PhyC without the SpyTag/SpyCatcher fusions was cloned by amplifying the PhyC ...
Western blots against the dynein HC were performed as above with the following modifications; lysates were diluted to 0.75 - 1mg/ml in Laemmli buffer, blocking buffer was 5% BSA in PBS-Tween (0.1%), membranes were probed with Rat dynein HC antibody (Santa Cruz Biotechnology) at 1:250 ...