Anti-Beta Catenin CTNNB1 Antibody Picoband™ (Monoclonal, 1F6)

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All paraffinized tissue blocks were cut at 4 μm thicknesses and detected by the SP immunochemistry kit (Zhongshan Golden Bridge Biotechnology, Beijing, China). IHC assay was conducted as previously described [27]. The rabbit monoclonal anti-human AJAP1 antibody (Bioss, China) at 1:100 dilution or the mouse monoclonal anti-human β-catenin antibody (CTNNB1, Boster) at 1:200 dilution was used for IHC. Two senior pathologists (Yun Niu and Shuhua Lv) evaluated the score without any knowledge of the clinicopathological outcomes of the patients. The percentage of positivity of the tumor was scored as “0” (no tumor cells), “1” (1–25%), “2” (26–50%), “3” (51–75%), and “4” (> 75%). The staining intensity of the positive tumor cells was scored as “0” (no staining), “1” (weak staining), “2” (moderate staining), and “3” (strong staining). As for AJAP1, the multiplier of the scoring of (0–3) for low expression and (4–12) for high expression were used. The IHC staining results of β-catenin were evaluated independently according to the subcellular localization of the nucleus and membrane. A positive/abnormal nuclear expression was defined as over 10% of the nuclear-stained tumor cells. An abnormal membranous expression demonstrated either no immunoreactivity in the membrane or less than 10% of the cancer cells with a positive membranous staining [28, 29].

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