Figure EV3. Validation of the reactivity of the anti‐PrP Fabs by Western blot, ELISA, and immunoprecipitation.

• A, B
  Western blot analysis to compare the reactivity of the indicated Fabs to mouse (m) recPrP_23‐231 and human (h) recPrP_23‐230 (A) and to full‐length and truncated PrP^C in BHs of various mouse lines. Actin was used as loading control (B).
• C
  ELISA to compare the efficiency of Fab83 and POM1 to detect and quantify wt and mutant PrP^C levels in CAD5‐Prnp ^−/− cells transfected either with wt or mutant PrP^C (deletion mutants PrPΔ23‐31 or PrPΔ23‐27; ${\text{PrP}}_{23-27}^{K\to A}$ with lysine residues 23, 24, and 27 replaced by alanine; ${\text{PrP}}_{23-27}^{\mathrm{Mirror}}$ with KKRPK exchanged to KPRKK). All concentrations (% relative to wtPrP^C) were determined by interpolating the ELISA signal to a standard curve of mouse recPrP_23‐231.

Data information: ELISA data were performed in triplicates. Data represent the mean ± sem. Two‐way ANOVA with Dunnett post hoc test: *P < 0.05; **P < 0.01; ****P < 0.0001. n = 3 technical replicates.