Ultraview dabv3 kit

Tissue was immersed in 10 % formalin for 24 h, washed in PBS buffer, dehydrated and embedded in paraffin wax. Sections of 3 μm were cut, deparaffinized and rehydrated. H&E-staining was performed for orientation. Sections for immunohistochemistry were stained using the Ventana Benchmark Xt® (Ventana Medical Systems, Tucson, USA). Demasking was done using the CC2 protocol for heat induced epitope retrieval. Cell conditioning solution (CC2, Ventana) is a low pH buffer solution which was added to the tissue sections. Sections were then heated to 94 °C for a total of 44 min with addition of further CC2 every four minutes. Primary antibody (AQP1, Alomone Labs, Jerusalem, Israel, cat. no: AQP-001) was diluted 1:1,000 in DAKO diluent and sections were incubated with this for 32 min. Reaction was visualized with the ultraview DAB v3 kit (Ventana). Nuclei were counterstained by hematoxylin and enhanced by bluing agent.