Mouse anti human cd68 monoclonal antibody

The hearts were fixed in 4% paraformaldehyde for 24 h and then placed in 30% sucrose PBS buffer before being embedded at the optimal cutting temperature and frozen. Approximately 5-μm-thick sections were prepared for hematoxylin and eosin (H&E) staining and immunohistochemical analyses. For morphometric lesion analysis, the sections were stained with Mayer’s H&E (Atom Scientific, Manchester, UK). Sections for CD68 staining were fixed by immersion in ice-cold acetone/methanol (1:1) for 3 min. These sections were incubated in a blocking solution with 3% goat serum in PBS for 1 h at room temperature and then with monoclonal mouse anti-human CD68 antibody (Dako, Glostrup, Denmark) overnight. The sections were then incubated with a secondary antibody for 1 h at room temperature; subsequently, they were mounted on a mounting medium with DAPI for fluorescence (Vector Laboratories; Burlingame, CA, USA), and imaged using a fluorescence microscope (Leica, Wetzlar, Germany). The lesion area and CD68 positive area were measured by taking the average of three sections by using ImageJ 1.52 quantification (National Institutes of Health, Bethesda, MD, USA), as defined previously [60 (link),61 (link)].