Caldas Immunohistochemistry Protocol
Tissue microarrays were prepared using duplicate 0.6mm cores extracted from formalin-fixed paraffin-embedded blocks containing material from patient tumors and xenografts. These were run using Leica’s Polymer Refine Detection System (DS9800) on their automated Bond – III platform. The HIER’s (sodium citrate and tris EDTA pre-treatments) are all run at 100°C. The de-waxing and re-hydration prior to IHC are done on the automated Leica ST5020, as is the post-IHC de-hydration and clearing. Finally, the mounting is done on Leica’s CV5030. The slides were reviewed by a pathologist.
|Mouse monoclonal (ER 6F11) anti-ER
|Mouse monoclonal (PgR 636) anti-PR
|Rabbit Monoclonal (4B5) anti-HER-2/neu