Biochemicals & reagents
1) McGowan et al. (1989), Inhibition of calpain in intact platelets by the thiol protease inhibitor E-64d; Biochem. Biophys. Res. Commun., 158 432 2) Wilcox and Mason (1992), Inhibition of cysteine proteinases in lysosomes and whole cells; Biochem. J., 285 495 3) Mizushima et al. (2010), Methods in mammalian autophagy research; Cell, 140 313
E-64d is a cell-permeable derivative of E-64c, an inhibitor of calpain and other cysteine proteases, such as papain, cathepsin B and cathepsin L (1,2). Addition of this compound to cell cultures should be done in serum-free media as esterases in serum will cleave the ethyl ester and reduce cell permeability using a typical working concentration is 0.5-10 µg/ml. It also inhibits degradation of autophagic cargo inside autolysosomes (3).