Mucin 5AC Antibody (45M1): sc-21701

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"Paraffin sections (4 µm) were used for histological staining. PAS staining was performed to evaluate goblet cell hyperplasia. IHC staining was performed by the peroxidase-labeled streptavidin-biotin technique as we described elsewhere. The sections were incubated overnight at 4°C in the presence of anti-IL-17A (R&D systems, Minneapolis, MN, USA) and anti-MUC5AC antibodies (Santa Cruz, Santa Cruz Biotech, CA, USA). Thereafter, each section was incubated with a secondary antibody and then with horseradish peroxidase-labeled streptavidin complex (Zhongshanjinqiao, Beijing, China). The distribution of peroxidase was revealed by incubating the sections in a solution containing 3% 3,3-diaminobenzidine tetrahydrochloride before being counterstained with hematoxylin and coverslipped. Negative control studies were performed by replacing the primary antibodies with normal IgG in appropriate concentrations.

The PAS staining and MUC5AC staining were semi-quantitatively scored on a scale of 0 to 4 (0, no staining; 1, mild; 2, medium; 3, severe; 4, extremely severe) in 10 randomly selected HPFs (×400 magnification) and then averaged. The number of IL-17A+ cells was determined by counting the positive-staining cells in 10 randomly selected HPFs (×400 magnification) and then averaged."

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