Ab37861

The paraffin sections of cancerous and benign lesions were baked for 2 h at 60°C, deparaffinized with dimethylbenzene and hydrated in gradient ethanol. Tissue antigen retrieval was performed using citrate sodium buffer (pH 6.0) at 95°C for 15 min and cooled at room temperature for 30 min. Endogenous peroxidase was blocked with methanol containing 3% H₂O₂ for 10 min, then the slides were treated with 5% normal goat serum (SL2; Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) for 30 min to limit non-specific binding. The anti-Axl rabbit polyclonal antibody (1:100; ab37861; Abcam, Cambridge, MA, USA) was overlaid on the sections and incubated overnight at 4°C. Following re-warming at room temperature for 1 h and three washes in PBS (AR0030; Wuhan Boster Biological Technology, Ltd., Wuhan, China), sections were incubated with horseradish peroxidase-labeled secondary antibody (1:500; SA00001-2; Wuhan Sanying Biotechnology, Wuhan, China) for 30 min at room temperature. Diaminobenzidine (DAB-0031; Fuzhou Maixin Biotech Co., Fuzhou, China) was used for antigen detection. Subsequently, the slides were counterstained with hematoxylin for 10 sec, dehydrated in gradient ethanol and mounted, and then viewed under an optical microscope. Negative controls used for comparison were incubated with phosphate buffer saline instead of the primary antibody.