Detection of endogenous Rfa1 protein in whole cell extract of S.cerevisiae. The extract of strain BY4741 (10 μg) was separated on SDS-PAGE 10-20% gradient gel. The anti-Rfa1 antibody was used at 1/2,000 dilution. As the second antibody, HRP-conjugated goat anti-rabbit IgG was used at 1/10,000 dilution