Fig. 1.

Evaluation of antibodies in relation to species specificity and S129 phosphorylation state. The species specificity of asyn antibodies used in the assay development (see Table 1) was tested using Western blot analysis by loading three different lysates – rat brain samples overexpressing h-asyn (AAV hasyn rat), uninjected wild-type rats (uninj. WT rat) or asyn knockout mouse brain tissue (asyn k.o. mouse) (a). For lysates 50 μg of protein was loaded. Actin is shown as a loading control. The specificity of antibodies for pS129 state was assessed using recombinant pS129 h-asyn protein or S129A h-asyn protein. Two additional lanes were loaded with either rat brain lysate overexpressing h-asyn (AAV hasyn rat) or asyn knockout mouse brain lysate (asyn k.o. mouse) to evaluate unspecific binding of antibodies on actual samples (b). For lysates 100 μg of protein per lane and for recombinant h-asyn 2 ng of protein were loaded