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Table 1 Characteristics for immunostaining and scoring

From: Presence of HER4 associates with increased sensitivity to Herceptin™ in patients with metastatic breast cancer

Protein HER2 EC phospho-HER2 (pY1248) p27Kip1 Oestrogen receptor α Progesterone receptor
Antibody Rabbit mAb Mouse mAb Mouse mAb Mouse mAb Mouse mAb
Origin Thermo Scientific Thermo Scientific Lab Vision Novocastra Novocastra
Clone SP3 PN2A, Ab-18 DCS-72.F6, Ab-1 6F11 1A6
Dilution of primary antibody 1:50 1:100 1:100 1:60 1:40
Staining pattern Membrane Nucleus and cytoplasm Nucleus
Epitope retrieval Heat induced, 10 mM citric acid buffer, pH 7.3 Heat induced, 10 mM sodium-citric acid pH 6.0 Heat induced, 10 mM citric acid buffer, pH 7.3
Blocking Endogenous peroxidase blocking
Primary antibody 30 min, room temperature Overnight, 4°C 30 min, room temperature 60 min, room temperature  
Detection system EnVision™ Dual Link System; DAB + chromogenic substrate iVIEW™ DAB Detection Kit  
Scoring in accordance with HercepTest guidelines Immunoreactive score Manufacturer's guidelines
  1. Anti-HER1 to 4 and anti-Ki-67 antibodies were applied as described previously [27].
  2. EC = antibody binds to extracellular domain; HER = human epidermal growth factor receptor; mAb = monoclonal antibody.