Table 3.
Summary of the primary antibodies used and antigen retrieval methods for immunohistochemical investigation of the cell populations present in feline ocular tissues with mycobacterial lesions.
| Target | Antibody | Source | Antigen retrieval | Primary antibody concentration or dilution |
|---|---|---|---|---|
| Calprotectin | Mouse monoclonal antihuman macrophages, clone MAC387 (MCA874G) | BioRad | Proteinase K (Dako) 20 minutes at RT |
1:800 (1.25 µg/ml) |
| Iba1 | Rabbit polyclonal anti-Iba1 (019-19741) | Wako | 0.01 M sodium citrate buffer, pH 6.0 Overnight at 60°C |
1:500 (1 µg/ml) |
| CD3 | Rabbit polyclonal antihuman CD3 (A0452) | Dako | Proteinase K (Dako) 20 minutes at RT |
1:50 (8 µg/ml) |
| Pax5 | Mouse monoclonal antihuman B-cell-specific activator protein, clone DAK-Pax5 (M7307) | Dako | 0.01 M sodium citrate buffer, pH 6.0 20 minutes at 121°C a |
1:50 (3.14 µg/ml) |
Abbreviation: RT, room temperature.
a
Antigen retrieval performed using the Antigen Retriever 2100 (Aptum Biologics Ltd, UK).