Pe conjugatedanti her2 antibody

Trastuzumab (Herceptin) was obtained from F. Hoffmann-La Roche Ltd (Basel, Switzerland). The antibodies for Western blot against EGFR, phospho (p)-EGFR (Tyr1068), HER2, p-HER2 (Tyr1248), HER3, p-HER3 (Tyr1289), AKT, p-AKT (Ser473), glyceraldehyde-3-phosphate dehydrogenase, and the corresponding secondary antibodies were purchased from Cell Signaling Technology (Boston, MA, USA). Phycoerythrin (PE)-conjugated anti-EGFR and anti-HER3 antibodies and fluorescein isothiocyanate-conjugated goat antihuman immunoglobulin (Ig) G (IgG) were from eBioscience (San Diego, CA, USA). The PE-conjugated anti-HER2 antibody was from BD, and HER3 was from Sino Biological Inc (Beijing, China). LMAb3, a novel anti-HER3 mAb (IgG1 k) was prepared in our laboratory. The reagents for electrophoresis and hybridization nitrocellulose filter membranes were obtained from Bio-Rad (Hercules, CA, USA). The BCA protein assay and enhanced chemiluminescent (ECL) reagents were from Thermo Scientific (Waltham, MA, USA). The cell culture medium Dulbecco's modified Eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from HyClone (Logan, Utah, USA). Human NRG1-b1/HRG1-b1 were purchased from R&D (Minneapolis, MN, USA), CCK-8 were purchased from Dojindo (Kumamoto, Japan), and agarose (cell culture grade) were purchased from Sigma-Aldrich (St. Louis, MO, USA).