Figure 1

Detection of disease-associated α-synuclein ( αS ^D ) in M83 mice homogenates from whole brain by ELISA. A. Anti α-syn antibodies immunoreactivity was tested on 4 symptomatic M83 mice from 11 to 16 months old, in comparison with 6 asymptomatic mice from 2 to 5 months old. B6C3H (genetic background of M83 mice) or B6 αS-null mice were used as additional controls. Sandwich ELISA with rabbit anti-αS polyclonal (capture)/Syn514, LB509, 4D6, 8A5 (reporters) and with clone 42 (capture)/anti-pSer129 αS (PSer129) (reporter) antibodies allows sick mice to be distinguished from asymptomatic M83 mice, whereas ELISA with anti-αS rabbit polyclonal (capture)/clone42 (reporter) does not. Error bars represent S.D. B. Determination of the sensitivity of detection of αS^D by ELISA. Two-fold dilutions of brain homogenates from a sick mouse were tested, and a positive signal was obtained for 12.5 μg brain equivalents, with both LB509 and PSer129 antibodies. Cut-offs for LB509 and PSer129 were visualized by a line in the color of the antibodies. C. Determination of the sensitivity of detection of αS^D by Western blot. 200 μg brain equivalents were necessary to detect αS^D in Western blot with PSer129 antibody. Molecular weight markers (in kDa) are indicated on the left of the blot.