Primary anti rat cd31 antibody

After 7 days, the rats were sacrificed with an i.p. injection of sodium pentobarbital. The skin scaffolds were excised and fixed in 4% formaldehyde for histological analysis. The fixed samples were embedded in paraffin. Paraffin sections were stained with hematoxylin and eosin (H&E, Sigma). For immunohistochemistry, the samples were stained with primary anti-rat CD31 antibody (1:500, Abcam), followed with HRP-labeled secondary antibody (1:2000, Abcam). After that the DAB was used as chromogenic substrate to locate the vessels. For immunofluorescence, the samples were stained with primary anti-rat CD31 antibody (1:500, Abcam), followed with Alexa-Fluor-488-labeled secondary antibody (1:2000, Abcam). Cell nucleus were dyed with 4,6-diamidino-2-phenylindole (DAPI) for 45 s. The graphs were taken and merged on an inverted microscope (Nikon E200).