TY - JOUR
T1 - Applicability of three anti-PrP peptide sera including staining of tonsils and brainstem of sheep with scrapie
AU - Garssen, G.J.
AU - van Keulen, L.J.M.
AU - Farquhar, C.F.
AU - Smits, M.A.
AU - Jacobs, J.G.
AU - Bossers, A.
AU - Meloen, R.H.
AU - Langeveld, J.P.M.
PY - 2000
Y1 - 2000
N2 - Three rabbit antibodies (R521, R505, R524) were produced, and raised to synthetic peptides corresponding to residues 94-105, 100-111, and 223-234, respectively, of the sheep prion protein (PrP). Epitope mapping analysis revealed the monospecific character of antisera R505 and R524. In addition to the amino acid sequence against which it was raised, R521 also recognized Other small epitopes. ELISA and radio-immunoprecipitation were used to assess the relative immunoreactivities of the antisera to the normal sheep prion protein (PrP(c)). Highest reactivity was found for R521, followed by R505 and R524. According to Western blot analysis, all three sera specifically reacted with the prion proteins PrP(sc) and PrP27-30, extracted from the brain stem of a scrapie-affected sheep. Yet, with R505 not all of the lower molecular weight deglycosylated forms could be detected. Contrary to the immunoreactivities found with the PrP(sc) and PrP27-30 isoforms, only R521 recognised PrP(c) from a healthy sheep. The usefulness of all three anti-peptide sera in the immunohistochemical detection of PrP(sc) in brain stem and tonsils of scrapie-affected sheep was demonstrated and compared with an established rabbit anti-PrP serum. (C) 2000 Wiley-Liss, Inc.
AB - Three rabbit antibodies (R521, R505, R524) were produced, and raised to synthetic peptides corresponding to residues 94-105, 100-111, and 223-234, respectively, of the sheep prion protein (PrP). Epitope mapping analysis revealed the monospecific character of antisera R505 and R524. In addition to the amino acid sequence against which it was raised, R521 also recognized Other small epitopes. ELISA and radio-immunoprecipitation were used to assess the relative immunoreactivities of the antisera to the normal sheep prion protein (PrP(c)). Highest reactivity was found for R521, followed by R505 and R524. According to Western blot analysis, all three sera specifically reacted with the prion proteins PrP(sc) and PrP27-30, extracted from the brain stem of a scrapie-affected sheep. Yet, with R505 not all of the lower molecular weight deglycosylated forms could be detected. Contrary to the immunoreactivities found with the PrP(sc) and PrP27-30 isoforms, only R521 recognised PrP(c) from a healthy sheep. The usefulness of all three anti-peptide sera in the immunohistochemical detection of PrP(sc) in brain stem and tonsils of scrapie-affected sheep was demonstrated and compared with an established rabbit anti-PrP serum. (C) 2000 Wiley-Liss, Inc.
KW - Anti-peptide antibodies
KW - Epitope mapping
KW - Prion proteins
KW - Scrapie
KW - Western blot/immunohistochemistry
U2 - 10.1002/1097-0029(20000701)50:1<32::AID-JEMT6>3.0.CO;2-Q
DO - 10.1002/1097-0029(20000701)50:1<32::AID-JEMT6>3.0.CO;2-Q
M3 - Article
SN - 1059-910X
VL - 50
SP - 32
EP - 39
JO - Microscopy Research and Technique
JF - Microscopy Research and Technique
IS - 1
ER -