Was performed on 3 or 5 μm formalin-fixed paraffin-embedded specimens that were float-mounted onto silanized glass slides. ERα was analyzed by a sandwich protocol (polyclonal first antibody HC-20, Santa Cruz Biotechnology. Dallas, Texas, USA; and goat anti-rabbit-immunoglobulin, peroxidase labeled, Dako/K4011). Sections were counterstained with hematoxylin. ERα-positive cells were identified by the presence of a brown stain over the nucleus and cytoplasm. Five regions were analyzed, and tumor samples were considered ERα-positive when 20 or more cells per field were positively labeled in the nucleus, in the cytoplasm or in both, and were found in at least three fields at 40X. Negative controls were obtained by using only the secondary antibody (Dako/K4011). The positive control was performed employing ovary tissue.
Zambrano-Estrada X., Landaverde-Quiroz B., Dueñas-Bocanegra A.A., De Paz-Campos M.A., Hernández-Alberto G., Solorio-Perusquia B., Trejo-Mandujano M., Pérez-Guerrero L., Delgado-González E., Anguiano B, & Aceves C. (2018). Molecular iodine/doxorubicin neoadjuvant treatment impair invasive capacity and attenuate side effect in canine mammary cancer. BMC Veterinary Research, 14, 87.
+ Open protocol