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Table 1 Immunohistochemical protocols and scoring for ER, PR, AR, HER2 and basal markers

From: Androgen receptor expression in a Sri Lankan patient cohort with early breast carcinoma

Marker

Supplier

Clone

Antigen retrieval

Primary dilution

Amplification step

Interpretation

Binarized as positive

Binarized as negative

ER

Dako

M7047

Clone ID5

Pressure cooker in citrate buffer

1:60

Universal secondary antibody

Envision

(Dako K5007)

ASCOCAP 2010 guideline [22]

> = 1% of tumour cells positive

< 1% of tumour cells positive

PR

Dako

M3569

Clone PgR636

Pressure cooker in citrate buffer

1:50

ASCOCAP 2010 guideline [22]

> = 1% of tumour cells positive

< 1% of tumour cells positive

HER2

Dako

A0485

Microwave in citrate buffer

1:600

ASCOCAP 2013 guideline [23]

HER2 = 3+

HER2 = 0,1

CK5/6

Dako

M7237

Clone D5/16B4

Pressure cooker in citrate buffer

1:25

Proportion Score

0 = No cytoplasmic staining

1-Positive cytoplasmic staining in 1–33% of cells

2-Positive cytoplasmic staining in 33–66% of cells

3-Positive staining in the cytoplasm in > 66% of cells

Intensity Score

0-No cytoplasmic staining

1-Weak cytoplasmic staining

2-Moderate cytoplasmic staining

3Strong cytoplasmic staining

Total score ≥ 2

Total score < 2

34βE12

Dako

N1553

Clone 34βE12

Microwave in Dako Target Retrieval Solution (S1700)

Undiluted

CK14

Cell Marque

LL002

Pressure cooker in citrate buffer

1:100

EGFR

Dako

M7239

Clone E30

Proteolytic digestion with Dako Proteinase K

(S3020)

1:25

Atkins [24] et al.

Any membrane staining above background, whether or not it is completely circumferential.

No membrane staining above background in any tumor cell

AR

Dako

M3562

Clone AR 441

Pressure cooker in citrate buffer at pH 6 for 7–8 min

1:50

ASCOCAP 2010 guideline on immunohistochemical testing of ER/PR [22]

> = 1% of tumour cells positive

< 1% of tumour cells positive