Immunofluorescence of colon issues.
Paraformaldehyde-fixed colon tissues were embedded in paraffin for
analysis of F4/80+ cell infiltration. Sections were
washed three times with PBS and then exposed to 3.0%
H2O2 for 1h to block endogenous
peroxidase activity. Subsequently, the sections were blocked with 3%
BSA for 30min at room temperature and then incubated at 4°C overnight
with anti-F4/80 (#GB11027, 1:500 per mouse, Servicebio). The next day,
sections were placed in PBS (PH7.4) and washed 3 times on a decolorizing
shaker for 5min each time. The corresponding secondary antibody was
added and incubated for 50min at room temperature and away from light.
Fluorescence microscopy to analyze the results (Zeiss Axioplan 2).