Table 2.
Overview of immunohistochemical antibodies and reagents.
| Target antigen | Manufacturer | Dilution and incubation of primary antibody | Detection method and chromogen |
|---|---|---|---|
| Formalin fixed tissue | |||
| Mouse anti-pig CD3 | Southern Biotech Clone PPT3 | 1:1000 o/n 4 °C | Ultravision One DABa |
| Goat anti-pig IgA | Nordic Biosite Lot no: A100-102A-16 | 1:4000 1 h RT | Vectastain Elite-ABC – Goat AECb |
| Goat anti-pig IgG | Nordic Biosite Lot no: A100-104A-12 | 1: 7000 1 h RT | Vectastain Elite-ABC – Goat AECb |
| Frozen tissue | |||
| Mouse anti-pig CD4 | Southern Biotech Clone 74-12-4 | 1:500 1 h RT | Ultravision AEC |
| Mouse anti-pig CD8α | BD Pharmingen Clone 295/33-25 | 1:500 1 h RT | Ultravision AEC |
All formalin fixed tissue was exposed to antigen retrieval by microwave boiling in aTEG buffer pH 9.0 for 2 × 5 min, bCitrate buffer pH 6.0 for 5 min, and blocked for endogenous peroxidase in a solution of hydrogen peroxide (0.6%) for 15 min. RT = room temperature.