Alk antibody

Genetic variants significantly expressed in NP-MCP patients were detected using ARSM-PCR. Then the protein expression of the ARSM-PCR-detected variant was validated using IHC. Rabbit anti-EGFR polyclonal antibody (Abcam, Cambridge, MA, USA), anti-KRAS antibody (Thermo Fisher Scientific, Rockford, IL, USA), rabbit monoclonal ALK antibody (Cell Signaling Technology, Danvers, MA, USA), anti-ROS1 rabbit monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA), RET antibody (EPR2871, 1:250) (Epitomics, Inc., Burlingame, CA), and HER2 antibody (Ventana Medical Systems, Tucson, AZ, USA) were used for immunohistochemistry analysis.

Immunohistochemical staining was performed on a Ventana BenchMark XT automated slide-processing system at Ventana Medical Systems. Immunohistochemical staining was performed using 2-μm sections of TMA blocks. All slides were stained with ALK antibody (clone D5F3 Cell Signaling Technology) diluted 1:50 using a Ventana Ultraview DAB detection kit in a Ventana BenchMark XT processor (Ventana Medical Systems, Inc, Tucson, AZ). Antigen retrieval was a standard automated process on the Ventana BenchMark XT at 37°C for 16 minutes. All immunohistochemical stains were evaluated independently by two pathologists. Specimens were scored 3+ if strong granular cytoplasmic brown staining in tumor cells was present (excluding non-tumor cells: alveolar macrophages, cells of neural origin, glandular epithelial staining), and the absence of cytoplasmic staining was classified a negative result as described previously [37 (link)].