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, 30Â min, RT |
PTEN | Rabbit mAb (138G6) | Cell Signaling Technology, Danvers, MA, USA | Citrate buffer pH 6.0a | 1:100, 40Â min, RT |
Phospho-AKT (Thr308) | Mouse mAb (18F3.H11) | Rockland Immunochemicals, Inc., Gilbertsville, PA, USA | None | 1:100, 1Â h, RT |
Phospho-mTOR (Ser2448) | Rabbit mAb (49F9) | Cell Signaling Technology, Danvers, MA, USA | Citrate buffer pH 6.0a | 1:100, 40Â 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, 1Â h, RT |