Table 2. IHC reagents, protocols and scoring systems.

Protein	Clone	Clonality	Source	Dilution	Antigen retrieval	Scoring system	Cutoff
ER	6F11/2	Mouse monoclonal	Novocastra	1 in 70	Citrate buffer pH6, 30 min	Allred	>2
HER2	c-erbB-2	Humanised monoclonal	Dako	1 in 250	Citrate buffer pH6, 40 min	Herceptest	⩾2*
BCL2	124	Mouse monoclonal	Dako	1 in 200	Tris-EDTA buffer pH9, 20 min	Modified H-score	>10%
MCM2	1B10	Mouse monoclonal	Novocastra	1 in 25	Citrate buffer pH6, 20 min	NA	NA

Abbreviation: ER=oestrogen receptor.

Allred Scoring System: Staining intensity score: 1=weak, 2=moderate, 3=strong; Proportion score: 1=<1%, 2=1–10%, 3=11–33%, 4=34–66%, 5=>66% Total score=Intensity score+proportion score=0–8.

Modified H-score (0–300)=intensity (0–3) × percentage of stained cells.

HercepTest: 0=No staining or weak staining in ⩽10% of cells, 1=weak incomplete membranous staining in >10% of cells, 2=moderate circumferential membranous staining in >10% of cells, 3=strong circumferential membranous staining in >10% of cells.