Immunohistochemistry (IHC) was performed on a Ventana Benchmark XT autostainer using the XT UltraView diaminobenzidine kit (Ventana Medical Systems, Oro Valley, AZ, USA). The primary antibodies were as follows: monoclonal anti-pERK1/2 antibody (rabbit anti-phospho-p44/42 MAPK [Thr202/Tyr204] clone 20G11, Cell Signaling Technology, Danvers, MA, USA), anti-pAKT (Santa Cruz, sc-135650, 1:30, heat mediated antigen retrieval) and anti-p53 (Novocastra, DO-7, 1:200, heat mediated antigen retrieval).
Xt ultraview diaminobenzidine kit
Manufactured by Roche
Sourced in United States
The XT UltraView diaminobenzidine (DAB) kit is a laboratory equipment product designed for use in immunohistochemistry (IHC) and immunocytochemistry (ICC) applications. It provides a chromogenic detection system for visualizing target antigens in tissue sections or cell samples.
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2 protocols using xt ultraview diaminobenzidine kit
1
Immunohistochemical Analysis of Signaling Pathways
2
Immunohistochemical Analysis of HER2/neu, HER3, and NRG1
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Immunohistochemistry was performed using 5 μm whole standard tissue sections of FFPE tumour samples. For the detection of HER2/neu, a prediluted anti-HER2/neu rabbit monoclonal antibody (clone 4B5; Ventana Medical Systems, Oro Valley, Arizona, USA) was used as the primary antibody. The staining was performed on a Ventana Benchmark XT autostainer using the XT UltraView diaminobenzidine kit (Ventana Medical Systems) following the manufacturer's protocols. Staining of HER3 and NRG1 was performed using the Vectastain ABC-Kit Elite Universal detection system (Vector Laboratories, Peterborough, UK). For HER3 immunohistochemistry, a monoclonal rabbit antibody was used as the primary antibody (ab93739; Abcam, Cambridge, UK). NRG1 staining was performed using a polyclonal human antibody (HPA010964; Atlas Antibodies, Stockholm, Sweden).
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