Ab96869

Paraffin-embedded MPM specimens were cut into 5 mm sections for immunohistochemistry analysis. Sections were deparaffinized in xylene and then rehydrated with ethanol. For antigen retrieval, the specimens were incubated in a microwave oven (95 °C) in ethylene diamine tetraacetic acid (EDTA) (ZLI-9068, Zhongshan Golden Bridge Bio-technology, China) for 15 min. After natural cooling, endogenous peroxidase activity was blocked with 3% hydrogen peroxide for 15 min at room temperature. Then, sections were washed with phosphate buffer saline (PBS) for 5 min. Nonspecific antibody binding was blocked with goat serum (ZLI-9022, Zhongshan Golden Bridge Biotechnology, China) for 10 min. Subsequently, the specimens were incubated with rabbit polyclonal anti-mesothelin antibody (1:200, ab96869, abcam, UK) at 4 °C for overnight. Rewarming at 37 °C and then washed with PBS, the specimens were incubated with goat anti-rabbit secondary antibody conjugated with horseradish peroxidase (HRP) (PV-9000, Zhongshan Golden Bridge Bio-technology, China). Counter-stained with hematoxylin, the sections were rinsed, dehydrated and covered.