Anti pecam 1 polyclonal antibody

The gastrocnemius muscles in paraffin were cut into 5-μm-thick sections for immunohistochemical staining. The sections were incubated overnight at 4 °C with the anti-PECAM-1 polyclonal antibody (Santa Cruz Biotechnology Inc., Dallas, TX, USA) diluted 1:500 and then stained using the Simple Stain Mouse System (Nichirei, Tokyo, Japan). Five fields from each section were randomly selected for the blind counting of capillary endothelial cells under a light microscope (× 200) by one skilled investigator to determine the capillary density.

The soleus muscles in paraffin were cut into 5-μm sections for immunohistochemical staining. The sections were incubated overnight at 4 °C with the primary antibody (anti-vWF polyclonal antibody; DAKO Japan, Tokyo, Japan) diluted 1:600 and subsequently stained using the Simplestain rat system (Nichirei, Tokyo, Japan) according to the manufacturer’s instructions. The negative control was performed by omitting the anti-vWF antibody.
The sections were also stained with the primary antibody (anti-PECAM-1 polyclonal antibody; Santa Cruz Biotechnology Inc., Dallas, TX, USA) diluted 1:200 and visualized using Alexa fluor 594 anti-rabbit secondary antibody (Invitrogen, Carlsbad, CA, USA). Nuclei were stained with 4’-6-diamidino-2-phenylindole (DAPI). The capillary to muscle ratio was counted under a fluorescence microscope (×200).