Specific antibodies

Tumors excised from the peritoneal cavity of 32 patients with epithelial ovarian cancer (stage III and IV) as well as fragments of tumor-free peritoneum from the same patients were included in the study. The age of patients ranged from 42 to 60 years. The tissues were fixed in 4% formalin and were then processed as described in^{11 (link)}. The cancer tissue was identified using standard H + E staining. Peritoneal mesothelium located near the cancer cells was identified based on positive stainings of the D2-40, Wt-1, HBME-1, and calretinin with specific antibodies (Dako, Santa Clara, CA). Antigen visualization was performed using an Envision Flex (Dako).

Whole-cell lysate samples prepared in Laemmli buffer were resolved in a 12% gradient SDS-PAGE gel, and proteins were transferred onto a polyvinylidene difluoride (PVDF) membrane (Bio-Rad). The membranes were blocked with Sea Block blocking buffer (Thermo Scientific) and probed with specific antibodies against HBV core amino acids 1 to 149 (Dako) or GAPDH (Thermo Scientific). Bound antibodies were revealed using IRDye secondary antibodies (Li-Cor). The immunoblot signals were visualized and quantified with the Li-Cor Odyssey system.