Detection of PrP^res in non-nervous tissues.

<p><b>A:</b> PrP^res from dura mater and frontal cortex (1∶24 dilution) from the present case is compared to PrP^res of the frontal cortex from sCJDMM1 (type 1) and sCJDMM2 (type 2). <b>B:</b> Two film exposures of immunoblots from non-nervous tissues compared with that of the frontal cortex (1∶10 dilution). Pituitary gland, adrenal gland and uterus are clearly positive while the bands in the remaining preparations are considered to be non specific. <b>C:</b> PrP^res from skin (double TH loading, equivalent to 4 mg of wet tissue) is barely detectable in TH (Lane 3) compared with frontal cortex TH, which is diluted 1∶4 (lane 1) or 1∶130 (lane 2). Skin PrP^res is better detectable along with the kidney PrP^res after sodium phosphotungstate (NaPTA) precipitation of PrP^res (Lanes 4 and 5), especially after long exposure (Lanes 4L and 5L) (kidney TH loaded in double amount; probed with mAb 3F4). <b>D:</b> PrP^res from mesenteric lymph nodes and other visceral organs recovered following NaPTA precipitation and compared with TH from the frontal cortex (1∶120 dilution) and sCJDMM1 following two film exposures. All organs but ascending colon are positive. Of note, unglycosylated isoform is underrepresented in all NaPTA precipitated samples as compared with that of the TH preparations (see panel A, lanes 3 and 4 and panel B). <b>A–D:</b> Membranes were probed with the mAb 3F4.