The relevance of the intrinsic subtype to the clinicopathological features and biomarkers in Japanese breast cancer patients

Table 1 Experimental conditions of immunohistochemistry assays

Antibody (clone)	Dilution	Source	Criteria for positive/negative
ER : Antibody (ID5)	×100	Dako	Positive: Stained cells >10%
PR : Antibody (PgR636)	×500	Dako	Positive: Stained cells >10%
HER2 (HercepTest)	Ready to use	Dako	HercepTest criteria
EGFR (EGFR pharmDx kit)	Ready to use	Dako	EGFR pharmDx kit criteria
CK5/6 (D5/16 B4)	×100	Dako	Positive: Stained cells >10%
PTEN (6H2.1)	1:100	Dako	- No staining
			+ Decreased staining intensity
			++ Equal or increased staining intensity
IGF-1R (C-20)	1:50	Santa Cruz	Score 0, 1+, 2+, 3+ using HercepTest criteria
Bcl-2 (124)	×50	Dako	Positive: Stained cells >10%
c-Kit (polyclonal; CD117)	×500	Dako	Positive: Stained cells >10%
c-Met (C-28, polyclonal)	1:100	IBL	Positive: Stained cells >10%
HIF-1α (H1alpha67)	×100	Novus Biologicals	Positive: Stained cells >10%
PDGFRA (C20, polyclonal)	1:200	Thermo	Positive: Stained cells >10%
Survivin (12C4)	×200	Novus Biologicals	Positive: Stained cells >20%
VEGFR2 (polyclonal; Flk-1(A-3))	×100	Santa Cruz	Positive: Stained cells >10%
VEGF-A (VG1)	Ready to use	Thermo	Positive: Stained cells >10%

ER: estrogen receptor, PR: progesterone receptor, HER2: human epidermal growth factor receptor type 2, EGFR: epidermal growth factor receptor, CK5/6: cytokeratin 5 or 6, PTEN: phosphatase and tensin homolog deleted on chromosome 10, IGF-1R: insulin-like growth factor-1 receptor, Bcl-2: B-cell lymphoma 2, c-Met: hepatocyte growth factor receptor, HIF-1α: hypoxia-inducible factor 1-alpha, PDGFRA: alpha-type platelet-derived growth factor receptor, VEGFR2: vascular endothelial growth factor receptor 2, VEGF-A: vascular endothelial growth factor A.
NOTE. Dako: Tokyo, Japan. Santa Cruz: Tokyo, Japan. IBL: Gunma, Japan. Novus Biologicals: Littleton, Colorado, USA. Thermo: Waltham, Massachusetts, USA.

ISSN: 1477-7819