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Table 1 Experimental conditions of immunohistochemistry assays

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

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%

  1. 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.
  2. NOTE. Dako: Tokyo, Japan. Santa Cruz: Tokyo, Japan. IBL: Gunma, Japan. Novus Biologicals: Littleton, Colorado, USA. Thermo: Waltham, Massachusetts, USA.