Detection of disease-associated α-synuclein ( αS
) 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 αSD 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 αSD by Western blot. 200 μg brain equivalents were necessary to detect αSD in Western blot with PSer129 antibody. Molecular weight markers (in kDa) are indicated on the left of the blot.