Multiplexed Immunohistochemistry for PD-L1 and Immune Cells
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Corresponding Organization : Dana-Farber Cancer Institute
Other organizations : Institut Gustave Roussy, Duke Medical Center, Memorial Sloan Kettering Cancer Center, Exelixis (United States)
Protocol cited in 1 other protocol
Variable analysis
- Antibody against PD-L1 (405. 9A11 mouse monoclonal antibody, 1:100, 13 micrograms/ml, Dr. Freeman laboratory, Dana-Farber Cancer Institute, Boston, MA, USA and commercially available through Cell Signaling Technology (CST))
- Cocktail of antibodies recognizing immune cells consisting of anti-CD45 (1:500, D9M8I XP, rabbit monoclonal antibody, CST) with anti-CD163 (1:5000, EPR19518, rabbit monoclonal antibody, Abcam)
- Presence and localization of PD-L1 and immune cells in the tumor sections
- Formalin-fixed and paraffin-embedded (FFPE) tissue sections
- Bond Rx Autostainer (Leica Biosystems, Buffalo Grove, IL)
- Bond Polymer Refine Detection Kit (DS9800; Leica Biosystems)
- Bond Polymer Refine Red Detection Kit (DS9390, Leica Biosystems)
- Bond Epitope Retrieval Solution 2 (EDTA, pH = 9.0) for 30 minutes
- Hematoxylin counterstaining, dehydration in graded ethanol and xylene, mounting, and cover slipping
- Not explicitly mentioned
- Not explicitly mentioned
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