VWF Antibody (C-12): sc-365712

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The whole ischemic limbs were harvested; the adhering tissues and femora were carefully removed, and samples were immersion-fixed with 4% buffered paraformaldehyde, performed on serial 5-μm-thick paraffin-embedded sections. Immunohistochemical staining was performed on mouse ischemic thigh muscle (sartorius muscle, gracilis muscle, adductor muscle and semimembranosus muscle were included) using goat anti-Von Willebrand factor (vWF; Santa Cruz Biotechnologies, Santa Cruz, CA, USA), rabbit anti-CD34 (Millipore, MA, USA), and goat anti-endothelial NO synthase antibodies (eNOS; Cell Signaling, CA, USA), followed by counterstaining with Hoechst. The stained slides were observed using fluorescence microscopy. Three cross-sections were analyzed for each animal, 10 different fields from each tissue preparation were randomly selected, and visible capillaries were counted. Capillary density is expressed as the capillary/myofiber ratio.

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