Immunohistochemical Analysis of HES1 Expression

Formalin-fixed paraffin-embedded specimens were processed for fine sections (5 μm) and were mounted on slides coated with poly-l-lysine (Sigma, St Louis, MO), followed by conventional hematoxylin and eosin staining for histologic assessment and immunohistochemistry (IHC). Citrate buffer (pH 6.0) was used for antigen retrieval treatment in the microwave and incubated overnight at 4°C with the primary rabbit polyclonal antibody of HES1 (ab87395; Abcam, Cambridge, United Kingdom) at a dilution of 1:250 respectively. An Envision System peroxidase kit (DAKO, Carpinteria, CA) was used for staining. Detailed methodology has been described earlier.^{19 (link)}