Quantitation of protein deposition by dot-blot ELISA. Glass slides were prepared as described in Fig 3. Immobilized protein was eluted by repeated scrape harvesting into a Tris-HCl buffer containing 10% SDS and applied to nitrocellulose membranes as described in Materials and Methods. Duplicate strips were allowed to react with antibodies against either vWF (upper row) or TSP (lower row). The weak signal generated by the anti-vWF antibody against TSP alone was less than 5% of the signal generated by reaction against the lowest concentration of vWF (1.25 μg/mL) (determined by densitometry). In separate experiments, neither the anti-vWF nor the anti-TSP antibody demonstrated detectable cross-reactivity against either TSP or vWF, respectively.