Los anticuerpos primarios utilizados, fueron: un anticuerpo policlonal anti-Met (Santa Cruz Biotechnology, CA, USA), un anticuerpo anti-HGF (Immuno-Biological Laboratories, Gumma, Japon) y un anticuerpo monoclonal anti-PCNA
(DAKO Corporation, CA, USA).
The sections were deparaffinized, submitted to antigen retrieval and incubated with the anti-p53, anti-PCNA
and anti-bcl-2 primary antibodies as shown in Table I.
Each staining run was based on either a specific monoclonal anti-Ki67 antibody (NCL-Ki67-MM1) or a specific monoclonal anti-PCNA
antibody (NCL-PCNA) for immunohistochemical staining of tumor specimens at each time points.
Sections were incubated overnight in a humid chamber at 4 [degrees] C with anti-PCNA
mouse antibody (monoclonal IgM; Beckman Coulter, Miami, FL) at a dilution of 1:500.
Sections were cut (4-6 [micro]m), mounted onto positively charged slides (Fisher-brand, Superfrost/Plus and ProbeOn Plus slides) and stained either with hematoxylin and eosin (H&E) (9) or with anti-PCNA
with a hematoxylin counterstain (Zymed, PCNA Staining Kit).