Evaluation of the duplex AlphaLISA performance. Assessment of the Acceptor-bead performance and comparison between Europium and Terbium based beads was carried out once for both the LB509 ( a ) and the 11A5 ( b ) antibodies. In both instances the biotinylated 4B12 antibody was used and signal to background ratio calculated against serial dilutions of h-asyn proteins spiked in naïve rat brain lysate. Presence of any hindrance of Acceptor-bead coupled antibodies against each other was assessed for LB509-Terbium ( c ) and 11A5-Europium beads ( d ). Cross-talk between the channels was assessed using the Resorufine/Amplex Red FP535 Terbium filter ( e ) and the Europium 615 filter ( f ). For these experiments the relevant analyte, Acceptor- and Donor-bead information is given under the x-axis. Both the hindrance of Acceptor-bead coupled antibodies and the channel cross talk were run twice. See Additional file 2 : Figure S2 for a theoretical explanation of the phenomena. AU arbitrary units
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