Ki 67 antibody

Four-micrometer-thick sections from FFPE tissue blocks were cut with a microtome and routinely deparaffinized. The sections were incubated with 0.3% hydrogen peroxide. The antigen retrieval procedure was performed in 0.01 M of citrate buffer (pH 6.0) or Tris-EDTA Buffer (10 mM Tris, 1 mM EDTA, 0.03% Tween 20 (pH 9.0)) at 95 °C, and counterstaining was conducted with hematoxylin. The STAT6 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-621, 1:400 dilution) was used for STAT6 immunohistochemical staining. The CD34 antibody (Thermo Fisher Scientific, Inc., MA1-22646, 1:100 dilution) and Ki-67 antibody (Novocastra, Buffalo Grove, IL, USA, NCL-Li-Ki-67-MM1, 1:50 dilution) were used. The immunohistochemistry (IHC) for APAF1 was performed using an anti-APAF1 antibody (Sigma, St. Louis, MO, USA, PRS2015, 1:400 dilution) and IHC for TP53 was performed using the TP53 antibody (Vector Laboratories, Burlingame, CA, USA, VP-P958, 1:50 dilution). Loss of immunohistochemical reactivity for APAF1 was defined as no (0) or weak (1+) staining intensity of the tumor cells. Positive immunohistochemical reactivity for TP53 was defined as near complete absence of the immunoreactivity or positive immunoreactivity in more than 50% of the tumor cells.