Rabbit anti-Transferrin (109-4134 lot 3033, green), Goat-anti-Alpha-1-Anti-Trypsin (100-101-147 lot 5842), and Mouse-a-GST (200-301-200 lot 24882) were used in a multiplex system to detect target proteins under reducing (R) conditions (+4% BME) in albumin depleted human serum with 320 ng of added GST. Sample was run by SDS-PAGE, transferred to 0.2 um PVDF using the BioRad Trans-Blot Turbo and blocked in 2.5% Blotto, 2.5% BSA, 0.02% Tween over night at 4°C. Membrane was probed with three primary antibodies at 1:1000 dilution (in MB-070 over night at 4°C). Detection shown was using DyLight549 Donkey anti-Rabbit IgG (611-742-127 lot 21100, shown as green) DyLight 488 Donkey anti-Mouse IgG (610-741-124 lot 21095, shown as blue), and DyLight 649 Donkey anti-Goat IgG (605-743-125 lot 20834, shown as red) at 1:10000 (in MB-070 30 min RT). Blots were washed, rinsed in methanol, dried and Images were collected using the BioRad VersaDoc System.