Fig. 3.
Western blotting analysis of PrP-res in tissue from sporadic CJD patients using the concentration method, which precipitates the sample by centrifugal separation.
The brain (1 mg) or tissue (100 mg of weight) homogenates, such as spleen, kidney, adrenal gland, liver, or lung from normal persons (a–b, upper panel) or sporadic CJD patient 2 (a, lower panel) and 4 (b, lower panel), were digested with PK and samples were prepared for Western blotting (see Materials and Methods). The tissues (100 mg) and brain (1, 10, or 100 μg) samples were loaded on SDS-PAGE, and PrPSc was detected using anti-PrP (3F4) monoclonal antibody and anti-mouse IgG antibody-conjugated HRP. The brain samples were used as a detection limit for Western blotting. The rate indicates measured band intensity, which was compared with brain samples. Recombinant human PrP is used as expose control. c) Detection limits were compared between concentration (C) and standard (S) method. PK-digested samples directly mixed with sample buffer in standard methods, which loaded to indicating weight.
(N.D.: Not detected.)