Immunohistochemical Profiling of ATM, pRAD50, and SLFN11
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Corresponding Organization : Institute of Cancer Research
Other organizations : Manchester Academic Health Science Centre, University of Manchester, The Christie NHS Foundation Trust, Institut Gustave Roussy, Inserm, Université Paris-Sud, Université Paris-Saclay, USC Norris Comprehensive Cancer Center, AstraZeneca (United Kingdom)
Variable analysis
- Antibody used for IHC (ATM, pRAD50, SLFN11)
- Expression level of ATM, pRAD50, and SLFN11 proteins (measured by H-score)
- Tissue type (formalin-fixed paraffin-embedded clinical tissues)
- Tissue section thickness (4- to 5-μm)
- Antigen retrieval method (pH9 ER2 for 25 minutes)
- Staining platform (Leica Bond RX)
- Protein blocking (SignalStain)
- Detection method (Polymer Refine Detection Kit)
- Counterstaining (hematoxylin)
- Slide digitalization (Aperio AT2 scanner)
- Scoring method (pathology H-scores)
- Scorer (single pathologist at 20× magnification)
- Appropriate control tissues
- Matched isotype controls
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