Rabbit anti mouse ucp1 antibody

Paraffin-embedded tissue sections in 5-µm thickness were stained with hematoxylin and eosin (H&E) using our standard protocol (Lim et al., 2014 (link); Iwamoto et al., 2015 (link); Seki et al., 2016 (link)). For UCP1 and CD31 staining, the paraffin-embedded tissue sections were incubated with a rabbit anti–mouse UCP1 antibody (1:200; Abcam) and a rat anti–mouse CD31 antibody (1:200), followed by staining with a mixture of secondary antibodies comprising an Alexa Fluor 555–labeled goat anti–rat antibody (1:200; Invitrogen) and an Alexa Fluor 649–labeled goat anti–rabbit antibody (1:400; Jackson ImmunoResearch Laboratories Inc.). Positive signals were captured with a fluorescence microscope equipped with a Nikon DS-QilMC camera. A rat anti–mouse VEGFR1 antibody (1:100 dilution in blocking buffer) was used for incubation at 4°C overnight. A secondary goat anti–rat Alexa 555 antibody (A21434; 1:200 dilution; Invitrogen) in blocking buffer was incubated at room temperature for 1 h. For lipid staining, Oil Red O staining was performed according to the BioVision manual. In brief, frozen liver sample was fixed with 4% paraformaldehyde for 30 min, washed with 60% isopropanol, and stained at 37°C with Oil Red O (O1391; Sigma-Aldrich) solution for 15 min. Nuclei were counterstained with hematoxylin for 10 min, mounted with glycerol, and examined under a light microscope.