Immunohistochemistry of FFPE Tissues

Immunohistochemistry was performed on formalin-fixed, paraffin-embedded (FFPE) tissue sections using the avidin–biotin complex method as described previously [16 (link)]. We used 4-μm-thick serial sections of each FFPE blocks with antibodies against the following: CD3 (PS1; 1:100) from Santa Cruz Biotechnology (Santa Cruz, CA, USA), CD4 (368; 1:100), and CD8 (4B11; 1:200) from Leica Microsystems (Newcastle-upon-Tyne, UK), and FOXP3 (42; 1:100) produced in-house [4 (link)]. Immunohistochemistry without the primary antibody was performed as a negative control.

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Ino Y., Oguro S., Yamazaki-Itoh R., Hori S., Shimada K, & Hiraoka N. (2019). Reliable evaluation of tumor-infiltrating lymphocytes in pancreatic cancer tissue biopsies. Oncotarget, 10(10), 1149-1159.

Publication 2019

CD3 (PS1; CD4 (368; CD8 (4B11;

Antibodies Antibody Avidin Biotin Embedded paraffin Formalin Immunohistochemistry

Corresponding Organization :

Other organizations : Tokyo National Hospital

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Variable analysis

independent variables

Antibodies used for immunohistochemistry: CD3, CD4, CD8, FOXP3

dependent variables

Presence and localization of CD3, CD4, CD8, and FOXP3 proteins in formalin-fixed, paraffin-embedded (FFPE) tissue sections

control variables

FFPE tissue sections
Avidin–biotin complex method for immunohistochemistry
4-μm-thick serial sections of each FFPE block

controls

Negative control: Immunohistochemistry without the primary antibody

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