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Table 2 Details of immunohistochemical procedures

From: Epidermal growth factor receptor pathway mutation and expression profiles in cervical squamous cell carcinoma: therapeutic implications

Antigen

Antibody (clone)

Supplier

Antigen retrieval

Antibody dilution, incubation time

EGFR

Mouse mAb (H11)

Dako North America Inc., Carpinteria, CA, USA

0.1% (w/v) trypsin in 40 mM CaCl2/TBS pH 7.8, 20 min, 37°C

1:100, 30 min RT then 4°C overnight

PIK3CA

Rabbit pAb (LS-B5363)

LifeSpan Biosciences, Inc., Seattle, WA, USA

Citrate buffer pH 6.0a

1:150, 30 min, RT

PTEN

Rabbit mAb (138G6)

Cell Signaling Technology, Danvers, MA, USA

Citrate buffer pH 6.0a

1:100, 40 min, RT

Phospho-AKT (Thr308)

Mouse mAb (18F3.H11)

Rockland Immunochemicals, Inc., Gilbertsville, PA, USA

None

1:100, 1 h, RT

Phospho-mTOR (Ser2448)

Rabbit mAb (49F9)

Cell Signaling Technology, Danvers, MA, USA

Citrate buffer pH 6.0a

1:100, 40 min, RT

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204)

Rabbit mAb (D13.14.4E)

Cell Signaling Technology, Danvers, MA, USA

Citrate buffer pH 6.0a

1:400, 1 h, RT

  1. aHeat treatment: 105°C for 20 min (Decloaking Chamber, Biocare Medical, Concord, CA), then 20 min cool down.