Electrophoresis and total protein staining of the 20 PMSA substrates prepared with mouse PrP variants with all naturally occurring amino acids at position 108.

Substrates were evaluated to confirm comparable rec-PrP concentrations prior to assessing their spontaneous misfolding capacity. Except for the L108C variant, which formed dimers (indicated by an asterisk) due to potential disulfide bridge formation, all other variants exhibited similar concentrations and were deemed suitable for PMSA. Despite efforts to concentrate the L108C substrate, dimer formation limited its preparation. The assay proceeded with the available substrate, acknowledging that while the lower concentration might underestimate misfolding propensity, enhanced misfolding would remain detectable as misfolding is not significantly affected within a certain concentration range [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1012890#ppat.1012890.ref001" target="_blank">1</a>]. MW: Molecular weight marker. (PDF)