Dako envision hrp polymer detection system

The percent of Ki67-positive cells counted in triplicates in high-power fields (400×) was used as a measure of proliferation. Briefly, formalin-fixed paraffin-embedded prostate sections were hydrated, subjected to antigen retrieval and incubated with anti-Ki67 antibody (M7240; Dako North America Inc., Carpinteria, CA). Slides were developed with using Dako Envision™ +/HRP polymer detection system (K4001; Dako North America Inc.) and visualized with 3,3'-diaminobenzidine chromagen followed by hematoxylin counterstain.

The percentages of Ki67 and cleaved caspase-3 positive cells counted in triplicate under a high power field (400X) were used as a measure of proliferation and apoptosis, respectively. Briefly, formalin-fixed paraffin-embedded mammary and MCF10AT1 tumor sections were hydrated, subjected to antigen retrieval, and incubated at room temperature with 1:100 anti-Ki67 antibody (M7240, Dako North America Inc, Carpinteria, CA) for 30 min and 1:100 cleaved-caspase-3 antibody (9661, Cell Signaling, Danvers, MA) for 1 h. Slides were developed using the Dako Envision™ +/HRP Polymer detection system (K4001, Dako North America Inc, Carpinteria, CA) and visualized with 3,3'-Diaminobenzidine (DAB) chromagen followed by hematoxylin counterstain. Staining was completed using a Dako Autostainer Plus.