Mouse Anti-PRNP Recombinant Antibody (clone POM2) (CAT#: PABL-310)

Figure 1 Competition ELISA and epitopes of N-terminal specific POMs.

Competition ELISA with pools of 10 sequential peptides (all left panels) revealed a broad area of PrP including the epitope of each antibody; for example P11–P20 and P21–P30 for POM2 and P31–P40 for POM3. Subsequently, competition with single peptides (all right panels) representing the ones included in the competing pool from the left panel, identified the exact epitopes of POM2, 3, 11, 12 and 14.

Polymenidou, M., Moos, R., Scott, M., Sigurdson, C., Shi, Y. Z., Yajima, B.,... & Bellon, A. (2008). The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes. PLoS One, 3(12), e3872.

Figure 2 Linear 12-meric peptides do not compete with binding of C-terminal POMs to PrP.

Polymenidou, M., Moos, R., Scott, M., Sigurdson, C., Shi, Y. Z., Yajima, B.,... & Bellon, A. (2008). The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes. PLoS One, 3(12), e3872.

Figure 3 Cross-species specificities and binding idiosyncrasies of C-terminal specific POMs.

Western blot analysis on PrP variants, with various amino acids substituted by cysteine as indicated. Blots were loaded with equal protein amounts and a silver stained gel serves as a loading control (first panel). Only mild divergences in the binding patterns of the various POMs were found here. The most striking one is that 6H4, POM1 and POM17 are binding weaker to rmPrP with cysteine substitutions at positions 140 and 145.

Polymenidou, M., Moos, R., Scott, M., Sigurdson, C., Shi, Y. Z., Yajima, B.,... & Bellon, A. (2008). The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes. PLoS One, 3(12), e3872.

Figure 4 Immunohistochemistry on mouse brain for staining of PrPᶜ.

Consecutive brain sections of Tg20, WT, or Prnpo/o mice focusing near the hippocampus area were incubated with the same concentration of each POM or IgG1 control antibody. The same HRP-conjugated anti-mouse antibody was used for development, allowing direct comparison of the performance of each antibody. POM1 and POM19 seemed to be the most sensitive antibodies in this assay, since they could readily detect PrPᶜ in the wild-type brains.

Polymenidou, M., Moos, R., Scott, M., Sigurdson, C., Shi, Y. Z., Yajima, B.,... & Bellon, A. (2008). The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes. PLoS One, 3(12), e3872.

Figure 5 Immunoprecipitation, peptide competition and specific elution of PrPᶜ and PrPᶜ with POMs.

Brain homogenates from healthy or terminally scrapie sick mice were incubated with POM1–7, covalently coupled to paramagnetic beads. All antibodies tested immunoprecipitated PrPᶜ very efficiently and POM2, 5 and 6 showed competent immunoprecipitation of PK-digested PrPˢᶜ. Plasminogen was used as a positive and non-specific IgG as a negative control.

Polymenidou, M., Moos, R., Scott, M., Sigurdson, C., Shi, Y. Z., Yajima, B.,... & Bellon, A. (2008). The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes. PLoS One, 3(12), e3872.