Axl antibody

Melanoma tissue and paracancerous tissue were collected and fixed in paraformaldehyde at room temperature, rinsed with PBS for 3 times, and embedded in paraffin and sectioned. The sections were dewaxed with xylene-ethanol solution, followed by sodium citrate buffer (pH 6.0) for antigen repair, and rinsed with PBS for 3 times. They were put in 30% hydrogen peroxide solution and reacted for 30 minutes in dark at room temperature. They were rinsed with PBS for 3 times. They were blocked with 3% BSA at room temperature for 20 min, then, Axl antibody (Abcam) or Akt antibody (Abcam) was added and incubated overnight in the refrigerator at 4°C. They were rinsed with PBS for 3 times, and HRP-labeled secondary antibody (Abcam) was added at the appropriate concentration and incubated at room temperature for 30 min. They were rinsed with PBS 3 times, 5 minutes each time. DAB chromo-developing solution (Solarbio®, Life Science) was added for staining for 2 min, and sections were rinsed with running water. The hematoxylin solution was redyed for 2 min and rinsed with PBS for 15 min. The slides were dehydrated in ethanol, then, 80% glycerin was added to the slides, and the cover glass was sealed. Finally, microscope observation was performed and photographs were taken (magnification: ×200).