Table 5. Antibodies used in the immunohistochemistry study.
| Cell type | Cell Marker | Type | Antigen retrieval method * | Positive tissue control | Dilution | IHC detection protocol ** | Supplier | Code | Isotype / Clone |
|---|---|---|---|---|---|---|---|---|---|
| T helper cells | CD4 | Rabbit monoclonal | H2 20/95°C | Tonsil | 1:100 | rabbit HRP | Cell Marque Lifescreen | CMC10431021 | IgG / SP35 |
| cytotoxic T cells | CD8 | Rabbit monoclonal | H2(40) | Tonsil | 1:500 | rabbit HRP | Cell Marque Lifescreen | CMC1083100 | IgG / SP16 |
| T regulatory | FOXP3 | Rabbit monoclonal | H2(60) | Tonsil | 1:200 | rabbit HRP | Acris Antibodies | AM21067PU-M | IgG / SP97 |
| macrophages | CD68 | Mouse monoclonal | H2(30) | Tonsil | 1:200 | refine HRP | Novocastra Laboratories | NCL-L-CD68 | IgG2a_Kappa / 514H12 |
| macrophages | CD163 | Mouse monoclonal | H2(30) | Tonsil | 1:750 | refine HRP | Serotec | MCA1853 | IgG1 / EDHu-1 |
| nucleolated cells | MHC-I | Rabbit monoclonal | H1(30) | Tonsil | 1:500 | rabbit HRP | Epitomics | 2307–1 | IgG / EPR1394Y |
* Leica Bond Retrieval Buffer ER1 (30 min. at 100°C); Retrieval Buffer ER2 (for 30, 40 or 60 min at 100°C); Retrieval-Buffer ER2 for 20 min at 95°C.
** Rabbit HRP = Bond Polymer Refine HRP Kit from Leica (DS9800) without Postprimary Antibody (Rabbit anti-Mouse); Refine HRP: same Leica kit with Postprimary Antibody (Rabbit anti-Mouse)