Figure 1: Western blotting evaluation of the newly purified monoclonal antibody against pfetin. A. SDS-PAGE/western blotting exhibited that the antibody distinguished pfetin positive (case 1-3) and negative (case 4-6) cases. The positive and negative pfetin expression of these cases were examined in our previous studies [11]. B. Western blotting with 2D-PAGE separation using previously purified antibody (upper panel), and newly purified one (lower panel). Western blotting was achieved according to our previous report [9]. 5 and 50 micrograms of proteins were separated by SDS-PAGE and 2D-PAGE, respectively. The first separation in 2D-PAGE was performed using 24 cm length IPG DryStrip gel (pI ranges between 4 and 7, GE), and the second separation was done by a home-made SDS-PAGE gel according to our previous report [9]. This study was approved by the ethical committee of National Cancer Center and Juntendo University.