Immunoblotting: Osteopontin antibody was used at a 1/1,000 dilution to detect Osteopontin by Western blot. In lane 2 reactivity is shown against 250 ng of Human osteopontin. Lane 3 shows reactivity of MMP-cleaved osteopontin. Lane 1 shows the position of molecular weight markers. Use a 1/10,000 dilution of HRP conjugated Goat anti-Rabbit IgG (R1454HRP) for detection.