Fig. 1.

Workflow for in vivo biotinylation of vascular antigens. Animals were first perfused with saline (PBS) to remove blood, followed by biotinylation using an isotonic solution of sulfo-NHS-biotin. Unreacted NHS-groups were quenched by perfusion with a Tris-HCl buffer (pH 7.4). All buffers were kept ice-cold and the perfusion times were kept as short as possible to minimize potential tissue damage and disruption. After biotinylation, multiple organs were harvested and preserved for histological analysis, or immediately homogenized and subjected to proteomics analysis