Mouse Anti-PRNP Recombinant Antibody (clone mAb 132) (CAT#: FAMAB-0275CQ)

Figure 1 Epitopes of PrP antibodies with more N-terminal linear epitopes were largely hidden in the folded conformation of PrPPᴿᴱˢ.

Purified PrPᴿᴱˢ from wild-type mice inoculated with the Chandler strain and with the 22L strain were used to examine the epitope accessibilities of the indicated more N-terminal PrP antibodies using indirect ELISA. The linear epitopes are shown in parentheses. Each of the epitopes was much less accessible in the folded conformation than after treatments with chaotropic 3 or 8 M GdnHCl solutions. OD450 readings with background subtracted are indicated as the means and SD from triplicate wells. Similar resultswere obtained in at least three independent experiments.

Saijo, E., Hughson, A. G., Raymond, G. J., Suzuki, A., Horiuchi, M., & Caughey, B. (2016). PrPSc-specific antibody reveals C-terminal conformational differences between prion strains. Journal of virology, 90(10), 4905-4913.

Figure 2 The PrPˢᶜ-specific C-terminal antibody 6H10 recognized the abnormal conformations of Me7 and 22L, but not Chandler, PrPᴿᴱˢ. The OD450 readings of MAbs 132, SAF-70, 8H4, and 94B4 after 8 M GdnHCl treatment showed that approximately equivalent amounts of PrPᴿᴱˢ were loaded into each well in ELISA plates.

(A) MAb 132 was used to normalize the OD450 reading. (B) The same data were normalized to MAb 8H4 readings. Twofold dilutions of PrPᴿᴱˢ were used to show that the OD450 readings were in a responsive range (the nine bars on the right in panels A and B). (C) Because PrPᴿᴱˢ was treated with PK and several possible PK cleavage sites are at the C terminus of PrP, the C-terminal linear-epitope antibody R1 was used to examine whether the C-terminal epitopes were still present in PrPᴿᴱˢ. OD450 readings with background sub_x0002_tracted are indicated as the means and SD from triplicate wells. *, P<0.05; **, P<0.01; ***, P<0.005; one-way ANOVA followed by the FDR (BenjaminiHochberg) test. Similar results were obtained in at least three independent experiments.

Saijo, E., Hughson, A. G., Raymond, G. J., Suzuki, A., Horiuchi, M., & Caughey, B. (2016). PrPSc-specific antibody reveals C-terminal conformational differences between prion strains. Journal of virology, 90(10), 4905-4913.

Figure 3 MAb 6H10 differed in its binding to the 22L and Chandler PrPᴿᴱˢ lacking GPI anchors and N-linked glycosylations.

Purified GPIneg Chandler and 22L PrPᴿᴱˢ were tested for epitope exposure of 6H10 using indirect ELISA. The OD450 readings of MAbs 132, SAF-70, 8H4, and 94B4 after 8 M GdnHCl treatment showed that equivalent amounts of PrPᴿᴱˢ were loaded in each well in the ELISA plates, and MAb 132 was used to normalize the OD450 readings. Twofold dilutions of PrPᴿᴱˢ were used to show that the OD450 readings were in a responsive range. OD450 readings with background subtracted are indicated as the means and SD from triplicate wells. *, P<0.05; **, P<0.01; ***, P<0.005; unpaired two-tailed Student's t test. Similar results were obtained in at least three independent experiments.

Saijo, E., Hughson, A. G., Raymond, G. J., Suzuki, A., Horiuchi, M., & Caughey, B. (2016). PrPSc-specific antibody reveals C-terminal conformational differences between prion strains. Journal of virology, 90(10), 4905-4913.