Immunofluorescence and immunohistochemistry

Knee joint sections (4 μm) were deparaffinized at 37 °C for 30 min and then hydrated with xylene, graded alcohol; after antigen retrieval was performed, blocked for 10 min with 3% H2O2 methanol solution at room temperature, sections were then incubated with anti CD31(1:500, abcam ab182981), anti-ADAMTS5(1:100, Bioss), and anti COL-II (1:100, Affinity AF0135), incubated overnight at 4 °C; 50 UL of Sheep anti-rabbit polymer was added for 20 min at room temperature;DAB was added for color development hematoxylin counterstaining for 10 min. The protein expression was observed under a light microscope, and three areas with high expression were taken and photographed for storage (all pictures were taken at × 400).

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