Anti siglec f

Immunohistochemistry was performed on thin (5 µm) sections of formalin-fixed and paraffin-embedded tissues. Intestines were extensively washed with cold PBS before fixation. Tissue sections were stained with Periodic Acid-Shiff. For immunohistochemistry, tissues were stained with the following antibodies (Santa Cruz Biotechnology, Dallas, TX, USA): anti-p100/p52 (dilution 1:100); anti-Siglec-F (dilution 1:100), and anti-CCL11 (Abcam, Cambridge, MA, USA, dilution 1:50) and secondary fluorescent Abs. Nuclei were counterstained with DAPI.

Histological sections (6 μm) were cut from formalin‐fixed paraffin‐embedded tissue blocks and were stained with hematoxylin–eosin (H&E) under standard conditions. Immunohistochemical staining was performed using the DAKO Envision system (DAKO, Carpinteria, CA, USA). Sections were immunostained with antibodies against UCP1 (Sigma‐Aldrich, St Louis, MO, USA). Peroxidase activity was detected with 3‐amino‐9‐ethyl carbazole. The adipocyte area in selected fat tissue sections was measured using iSolution DT 36 software (Carl Zeiss, Oberkochen, Germany).
For immunofluorescence staining, frozen sections were stained with anti‐Siglec‐F (Abcam, Cambridge, UK) or anti‐CD206 (Abcam) antibodies at 4°C overnight. Immunofluorescence analyses were performed to identify macrophages and eosinophils by staining tissues with a combination of anti‐perilipin (Fitzgerald, MA, USA) with either anti‐F4/80 or anti‐Siglec‐F antibodies, respectively. After incubation with the corresponding fluorochrome‐conjugated secondary antibodies, the sections were mounted and visualized using an LSM510 confocal laser scanning microscope (Carl Zeiss) installed in the Center for University‐Wide Research Facilities (CURF) at Chonbuk National University.