Detection of endonenous Tfa1 in whole cell extract of S. cerevisiae by Western blotting, using the anti-Tfa1 antibody The antibody was used at 1/2,000 dilution. As second antibody, HRP-conjugated goat anti-rabbit IgG was used at 1/10,000 The apparent molecular mass (~65 kDa) is larger than calculated mass of 55 kDa, which may be due to its highly acidic cluster at C-terminal 106 amino-acids (Asp/Glu rich).