Expression, purification and gelatinolytic activity of rTsATG4B. A Expression and purification of rTsATG4B. Lane M: standard marker; lanes 1/2, lysate of BL21 bacteria harbouring pQE-80L/TsATG4B without/with IPTG induction; lane 3: purified recombinant TsATG4B; B Proteolytic activity of refolded TsATG4B as detected by gelatine zymography. Lane M: protein molecular weight marker; lane 1: transparent strips due to the proteolytic activity of refolded TsATG4B in acrylamide-gelatine gels; lane 2: refolded rTsATG4B pre-incubated with the protease inhibitor E-64; C Identification of refolded rTsATG4B by Western blotting. Lane M: protein molecular weight marker; lane 1: refolded rTsATG4B recognized by anti-TsATG4B serum (1:100 dilution); lane 2: denatured rTsATG4B recognized by anti-TsATG4B serum (1:100 dilution).