Immunohistochemistry panel segregates molecular types of hepatocellular carcinoma in Brazilian autopsy cases

Table 1 Antibodies and immunohistochemistry parameters used in this study

Antibody	Manufacturer	Clone	Species	Dilution	Retrieval1	Positive control	Staining pattern
EGFR PharmDx	Dako	2-18C9	Mouse	Pre-diluted	Proteinase K	Provided by manufacturer	Membrane
Ki67 Ag	Dako	MIB-1	Mouse	1:400	Standard	Tonsil	Nucleus
Caspase 3	DBS	3C SP03	Mouse	1:400	Standard	Tonsil/stomach	Cytoplasm
ERK1	DBS	Polyclonal	Rabbit	1:100	Standard	Breast carcinoma	Cytoplasm
ERK2	DBS	Polyclonal	Rabbit	1:400	Standard	Breast carcinoma	Cytoplasm
MET	Cell signaling	Polyclonal	Rabbit	1:50	Standard	Breast carcinoma	Cytoplasm
mTOR	Cell signaling	Polyclonal	Rabbit	1:50	Standard	Lymphoid tissue	Cytoplasm
K19	Novocastra	b170	Mouse	1:300	Standard	Cholangio-carcinoma	Cytoplasm
Vimentin	Dako	Vim3B4	Mouse	1:3000	Standard	Kidney	Cytoplasm
p53	Dako	DO7	Mouse	1:100	Standard	Breast carcinoma	Nucleus
Beta-catenin	BD	14Beta-catenin	Mouse	1:800	Standard	Tonsil	Membrane/nucleus
Cyclin D1	Dako	SP4	Rabbit	1:100	Steamer 10 mmol/L TRIS, 1 mmol/L EDTA, pH 9	Tonsil/intestine	Nucleus

¹Antigen retrieval was performed by steam heating for 40 min in 10 mmol/L citrate buffer, pH 6. BD: BD biosciences™; DBS: Diagnostic bioSystems™; IHC: Immunohistochemistry; K19: Keratin 19; mTOR; Mechanistic target of rapamycin.