Tumor sections were evaluated for PD-L1 expression by IHC as reported previously (11 (link)). Briefly, 4–5-μm tumor sections were treated with 3% H2O2 for 10 minutes, blocked with 5% goat serum for 1 hour, and then incubated with a primary anti-human PD-L1 antibody (#13684, Cell Signaling Technology) at 1:500 dilution at 4°C overnight. Subsequently, using Dako CSAII Biotin-free Tyramide Signal Amplification System Kit, slides were incubated with secondary antibody and amplification reagent. Final staining was carried out by adding 3,3'-Diaminobenzidine.
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Immunohistochemical Evaluation of PD-L1
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Corresponding Organization : Sidney Kimmel Comprehensive Cancer Center
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Variable analysis
independent variables
- PD-L1 antibody concentration (1:500 dilution)
- PD-L1 expression in tumor sections
- Tumor section thickness (4-5 μm)
- Treatment with 3% H2O2 for 10 minutes
- Blocking with 5% goat serum for 1 hour
- Incubation time with primary antibody (overnight at 4°C)
- Dako CSAII Biotin-free Tyramide Signal Amplification System Kit
- Addition of secondary antibody and amplification reagent
- Final staining with 3,3'-Diaminobenzidine
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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