The PDAC tissues were incubated with anti-RHAMM IgG (Abcam, Cambridge, MA, USA) at a 1:100 dilution for 2 h at room temperature. The sections were incubated for 10 min with biotinylated anti-rabbit antibody for RHAMM according to the manufacture's instruction of Histofine SAB-PO (R) kit (Nichirei Biosciences Inc, Tokyo, Japan). The positive control was archival tissue from a normal human testis. The scoring method, which combined intensity and the percentage of positivity, was previously described 9 (link). The slides were scored by two individual researchers (N.S. and X.C.) on a blinded basis. The extent and intensity measures for each score were combined to identify “weak” expression or “strong” expression.
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Immunohistochemical Analysis of RHAMM Expression
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Corresponding Organization : Fourth Affiliated Hospital of China Medical University
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Variable analysis
independent variables
- Anti-RHAMM IgG (Abcam, Cambridge, MA, USA) at a 1:100 dilution
- RHAMM expression
- Incubation time (2 h at room temperature)
- Biotinylated anti-rabbit antibody for RHAMM (Histofine SAB-PO (R) kit, Nichirei Biosciences Inc, Tokyo, Japan)
- Incubation time with biotinylated antibody (10 min)
- Archival tissue from a normal human testis
- Combined intensity and percentage of positivity, as previously described
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