Alteration of somatostatin receptor 2 expression in canine mammary gland tumor

SAKAI, Kosei; YONEZAWA, Tomohiro; YAMAWAKI, Hideyuki; OYAMADA, Toshifumi

For Western blotting, 12 canine mammary gland tissues (4 normal, 4 simple adenomas, one complex adenoma and 3 simple carcinomas), harvested and flash frozen in liquid nitrogen, were obtained either from experimental animals or from patients undergoing surgery for mammary gland tumors at the Kitasato University Veterinary Teaching Hospital. We performed Western blotting analysis according to a previously reported protocol [12] . Twelve micrograms of protein were loaded in each well. The human SSTR2 antibody used in immunohistochemistry analysis (1:300 dilution) was also used in Western blotting. Samples were visualized using horseradish peroxidase-conjugated secondary antibodies and the EZ-ECL system (Biological Industries, Kibbutz Beit Hesmek, Israel). The expression level of SSTR2 in each sample was normalized to total actin (1:500 dilution, MAB1501, Sigma-Aldrich, St. Louis, MO, U.S.A.) expression on the same membrane as an internal control.