Microplates Microlon/F-shape REF 655092 (Greiner) were coated with 1 μg/mL of human 3D6 [29] overnight at room temperature in coating buffer (sodium carbonate pH = 9.6 0.05 M NaCO3 in MQ water). After washing plates (washing buffer 0.05% Tween- 20 in PBS were blocked with 4% not fat dry milk and incubated with brain homogenates or with Aβ to generate the standard curve. Next plates were washed, incubated with 50 ng/mL of biotinylated human 20C2 [29], and washed again. Finally, plates were incubated with streptavidin-HRP (Jackson ImmunoResearch Laboratories) diluted 1:8000 and developed using 3,3′,5,5′-Tetramethylbenzidine (TMB). The absorption was measured at 450 nm within 30 min of stopping the reaction with 2 M H2SO4 using the Perkin Elmer 2030 manager system.

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