Immunohistochemistry methods
| Protein | Section | Antigen retrieval | Primary antibody |
|---|---|---|---|
| For the Fas staining human tonsil was used as the positive control, for Bcl-xl and Bax staining Reed-Sternberg cells were used as the positive control. | |||
| COX-2, cyclooxygenase 2; iNOS, inducible nitric oxide synthase; MW, microwave at 700 W; SDS, sodium dodecyl sulfate. | |||
| Fas | Paraffin wax | MW 3×10 min at 98°C in 1mM EDTA; pH 8.0 | Mouse monoclonal at 1/400 dilution; Upstate Biotechnology, Lake Placid, USA |
| Bcl-xl | Paraffin wax | MW 2×15 min at 98°C in 0.1M Tris HCl; pH 9.0 | Mouse monoclonal at 1/100 dilution; Zymed Laboratories, South San Francisco, California, USA |
| Bax | Paraffin wax | MW 1×8 min boiling in 10mM citrate; pH 6.0 | Mouse monoclonal at 1/400 dilution; Santa Cruz Biotechnology P19, Santa Cruz, California, USA |
| Bcl-2 | Paraffin wax | MW 1×8 min boiling in 10mM citrate; pH 6.0 | Mouse monoclonal at 1/50 dilution; Dako, Glostrup, Denmark |
| COX-2 | Paraffin wax | MW 3×10 min at 98°C in 1mM EDTA; pH 8.0 | Mouse monoclonal at 1/50 dilution; Transduction Laboratories, Lexington, Kentucky, USA |
| iNOS | Paraffin wax | MW 3×10 min at 98°C in 1mM EDTA; pH 8.0 | Mouse monoclonal at 1/50 dilution; Transduction Laboratories |
| Active caspase 3 | Paraffin wax | MW; 1×8 min in 10mM citrate buffer; pH 6.0 | Rabbit polyclonal at 1/100 dilution; Cell Signalling Technology, Beverly Massachusetts, USA |
| Ki-67 | Paraffin wax | 3×15 min at 115°C in 0.2% SDS in maleate buffer; pH 6.0 | Mouse monoclonal MIB-1 at 1/400 dilution; Immunotech, Marseille, France |