Cell lysates of 293T and HeLa (20ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human PPIF (1:500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

Paraffin embedded sections of human breast cancer tissue were incubated with anti-human PPIF (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB)