Table 2. Primary antibody protocols.
Details of the primary antibodies and the antigen retrieval method used to detect the basement membrane (Col IV); terminal epidermal differentiation (Loricrin); migration (Vimentin); proliferation (Ki-67); and invasion (S100).
| Antibody | Primary antibody | Antigen retrieval method | ||||
|---|---|---|---|---|---|---|
| Antibody type | Source | Dilution | Time and temperature | Buffer | Time and temperature | |
| Collagen IV (Col IV) | Mouse | DKSH, Australia | 1:50 | 1 h, 37°C | Sodium Citrate (pH6.0) | 20 min, 80°C |
| Ki-67 | Mouse | Sigma Aldrich, Australia | 1:100 | 1 h, 37°C | EDTA (pH8.0) | 30 min, 90°C |
| Loricrin | Rabbit | Dako, Australia | 1:100 | 1 h, 37°C | EDTA (pH8.0) | 5 min, 97°C |
| S100 | Rabbit | Dako, Australia | 1:3,000 | 1 h, 37°C | Sodium Citrate (pH6.0) | 5 min, 95°C |
| Vimentin | Rabbit | Thermo Scientific, Australia | 1:800 | 12 –24 h,4°C | Sodium Citrate (pH8.0) | 20 min, 80°C |