VHIO Immunohistochemistry Protocol

Use standard immunohistochemistry procedures and the following unmasking conditions and primary antibody dilution for each antibody:

HER2

• Microwave in citrate buffer pH 6 20min.
• Primary antibody: c-erbB2 1/500 30min RT (A0485 Agilent).

ER

• Microwave in Tris- EDTA Buffer ph 9 20 min.
• Primary antibody: ER non-diluted 2h RT or o/n 4ºC (IS657, Dako)

Ki67

• Microwave in citrate buffer pH 6 20min.
• Primary antibody: ki67 1/100 60min RT (M7240, Dako)