Immunofluorescent Analysis of CD204 and CD206

Sections were deparaffinized and hydrated before antigen retrieval in 10 mM citric acid buffer. Then, sections were incubated in 1% Triton X-100 for 15 min. After eliminating endogenous peroxidase activity with 3% hydrogen peroxide for 15 min, sections were rinsed with PBS and incubated with primary anti-CD204 (cat. no. ab123946; 1 : 50; Abcam Inc.) and anti-CD206 (cat. no. ab125028; 1 : 50; Abcam Inc.) at 4°C overnight. After that, the sections were washed with PBS and incubated with specific secondary antibodies for 30 min at room temperature. Sections were then washed with PBS and sealed with Fluoromount-G™ Slide Mounting Medium (SouthernBiotech, Birmingham, AL, USA) as described before [22 (link)]. Images were taken using a fluorescent microscope and were analyzed using an Image Analysis system, version 11.0.